Toluidine sulfonamides and their use

ABSTRACT

The present invention provides novel compounds that inhibit cell proliferation and cell division and that inhibit the activation of Hypoxia Inducible Factor (HIF)-mediated transcription and signaling under hypoxic conditions. In one aspect, the compounds of the present invention are useful for the preparation of a medicament for the treatment or prevention of a disease or disorder selected from the group consisting of an inflammatory disease, a hyperproliferative disease or disorder, a hypoxia-related pathology and a disease characterized by excessive vascularisation. Also provided is a pharmaceutical composition comprising a compound of the invention and a second therapeutic agent or radiation useful for the treatment or prevention of the mentioned diseases or disorders. In a first aspect the present invention relates to a compound having a structure according to formula (I).

The present invention provides novel compounds that inhibit cellproliferation and cell division and that inhibit the activation ofHypoxia Inducible Factor (HIF)-mediated transcription and signalingunder hypoxic conditions. In one aspect, the compounds of the presentinvention are useful for the preparation of a medicament for thetreatment or prevention of a disease or disorder selected from the groupconsisting of an inflammatory disease, a hyperproliferative disease ordisorder, a hypoxia-related pathology and a disease characterized byexcessive vascularisation. Also provided is a pharmaceuticalcomposition, comprising a compound of the invention and a secondtherapeutic agent or radiation, useful for the treatment or preventionof the mentioned diseases or disorders.

BACKGROUND OF THE INVENTION

The normal response of cells to inadequate oxygen supply is mediated bythe hypoxia signaling pathway. This response is important for a numberof physiolocial functions such as tumor development and metastasis,resistance to apoptosis, induction of new blood vessel formation, andmetabolism among others. For a general review on hypoxia signaling seee.g. Qingdong Ke and Max Costa, Molecular Pharmacology (2006), vol. 70,no. 5.

As a result of hypoxia, augmented levels of a heterodimeric complex oftranscription factors (Hypoxia Inducible Factor, HIF), most notablyHIF-1α and HIF-1β, are observed in e.g. tumors to compensate incooperation with additional co-factors for the reduced availability ofoxygen and nutrients in this fast growing tissue type. Under anaerobicconditions, homeostasis of HIF-1α is imbalanced by its reduceddegradation, thus enabeling enhanced signaling through the HypoxiaResponsive Element (HRE) and resulting in increased expression of alarge number of survival and growth factors.

Hypoxic conditions are also found in non-tumor tissue. For example,retinopathy is a general term that refers to non-inflammatory damage tothe retina of the eye. This condition is most commonly caused by aninsufficient blood supply leading to hypoxia. Particularly people withdiabetes mellitus are at risk of retinopathy. The lack of oxygen in theretina of diabetics causes fragile, new, blood vessels to grow along theretina and in the clear, gel-like vitreous humour that fills the insideof the eye. Without timely treatment, these new blood vessels can bleed,cloud vision, and destroy the retina. Fibrovascular proliferation canalso cause fractional retinal detachment. The new blood vessels can alsogrow into the anterior chamber of the eye and cause neovascularglaucoma.

Recently, evidence has accumulated that inhibition of HIF-1 activitycould also act to prevent inflammation, by virtue of its role in theactivation and infiltration of macrophages and neutrophils into affectedtissues (see e.g. Giaccia et al., Drug Discovery, vol. 2, October 2003).

For the above-outlined reasons, compounds that inhibit HIF function arevaluable medicaments for the treatment or prevention of a disease ordisorder selected from the group consisting of an inflammatory disease,a hyperproliferative disease or disorder, a hypoxia-related pathologyand a disease characterized by excessive vascularisation.

Because of the importance of HIF-1 in tumor development, progression andmetastasis, a considerable amount of effort has been devoted to identifyHIF-1 inhibitors for cancer therapy. A number of small molecules and RNAconstructs, like siRNA, have been reported to exhibit inhibition of theHIF-1 pathway, e.g. Kung A L et al, Cancer Cell (2004), vol. 6, p. 33ff; Rapisarda A, et al. Cancer Res. (2002), vol. 62, p. 4316 ff.; Tan C.et al, Cancer Res. (2005), vol. 65, p. 605 ff; Mabjeesh N J, et al,Cancer Cell, (2003), vol. 3, p. 363ff; Kong X, et al, Mol Cell Biol(2006), vol. 26, p. 2019 ff.; Kong D, et al, Cancer Res. (2005), vol.65, p. 9047 ff.; Chau N. et al., Cancer Res. (2005), vol. 65, p. 4918ff; Welsh S, et al., Mol Cancer Ther (2004), vol. 3, p. 233 ff. However,these compounds often have activities other than HIF-1 inhibition, andmost of them lack the desired pharmacokinetic properties or toxicityprofiles required for a useful pharmaceutical agent. Furthermore, someof the compounds have the disadvantage that they can not be administeredorally, such as the HIF-1 inhibitor EZN-2968, which is a locked nucleicacid antisense oligonucleotide.

The scientific literature cited above emphasizes the high medical needfor new therapeutic agents to provide more efficient treatment ofdifferent proliferative and inflammatory diseases or disorders,hypoxia-related pathologies and diseases characterized by excessivevascularisation.

SUMMARY OF THE INVENTION

The present invention provides novelcompounds capable of prevention ortreatment of a disease or disorder. Data presented herein establish thatcompounds according to the present invention are surprisingly verypotent inhibitors of (i) the activation of HIF mediated transcriptionunder hypoxic conditions, of (ii) cell cycle and cell proliferation, of(iii) estrogen response element—mediated transcriptional activity, and(iv) that these compounds induce apopotosis.

In a first aspect the present invention relates to a compound having astructure according to formula I:

wherein

-   R¹ is selected from a group consisting of H, alkyl, alkenyl,    alkynyl, —CN, halogen, —OH, alkoxy, —SH, S-alkyl, —NH₂, NH-alkyl,    N-bis-alkyl, NHOH, NMeOH, NMe(OMe), —NO₂, —CF₃, —OCF₃ and C₁-C₄    hydroxyalkyl.-   R² is H or C₁-C₄ alkyl;-   R³ is H or —CH₃;-   R⁴ is phenyl or monocyclic 5- or 6-membered heteroaryl; optionally    substituted with one or more substituents selected from the group    consisting of:    -   alkyl, alkenyl, alkynyl, alkoxy, halogen, —CN, —CF₃, —OCF₃,        C₁-C₄ hydroxyalkyl, —OH, —SH, S-alkyl, —CN, N-bis-alkyl,        cyanoacetylene, —NO₂, —NR⁷R⁸, —C(O)R²⁰, N—O (wherein the        nitrogen atom is integral part of the monocyclic 5- or        6-membered heteroaryl)    -   and two substituents which form together a dioxymethylene bridge        (—O—CH₂—o—);-   R⁵ is H or —CH₃;-   R⁶ is selected from the group consisting of H, halogen, alkyl,    alkoxy, alkenyl, alkynyl, S-alkyl, —OH, —NR⁷R⁸, —CN, N-bis-alkyl,    —SH, —CF₃ and —OCF₃; or R⁶ forms together with R¹ a dioxymethylene    bridge (—O—CH₂—O—);-   R⁷ is H or alkyl;-   R⁸ is H or C₁-C₄ alkyl; and-   R²⁰ is C₁-C₄ alkyl;    with the proviso that R⁴ is not 3-alkoxy-pyridazine-5-yl; that if R⁴    is phenyl, then the 2- and 5-position of the phenyl ring may not be    substituted with two methoxy substituents at the same time; and that    R³ and R⁵ are not at the same time H.

In a further aspect the present invention relates to a pharmaceuticalcomposition comprising a compound according to the invention orpharmaceutically acceptable salt thereof and a second therapeutic agentuseful for the treatment or prevention of a disease or disorder selectedfrom the group consisting of an inflammatory disease, ahyperproliferative disease or disorder, a hypoxia related pathology anda disease characterized by pathophysiological hyper-vascularisation,and, optionally, a pharmaceutically acceptable carrier or excipient.

In a further aspect the present invention is directed at the use of acompound according to the invention or the composition according to theinvention for the preparation of a medicament for the treatment orprevention of a disease or disorder selected from the group consistingof an inflammatory disease, a hyperproliferative disease or disorder, ahypoxia related pathology and a disease characterized bypathophysiological hyper-vascularisation.

In another aspect, the invention provides a method for treating ahyperproliferative disease or disorder comprising administering acompound or composition according to the invention to a patient priorto, during and/or after he was subjected to a radiation therapy, achemotherapy, an immunotherapy, a laser/microwave thermotherapy or agene therapy using antisense DNA and RNA.

DETAILED DESCRIPTION

It is to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto limit the scope of the present invention which will be limited onlyby the appended claims. Unless defined otherwise, all technical andscientific terms used herein have the same meanings as commonlyunderstood by one of ordinary skill in the art.

Preferably, the terms used herein are defined as described in “Amultilingual glossary of biotechnological terms: (IUPACRecommendations)”, Leuenberger, H. G. W, Nagel, B. and Klbl, H. eds.(1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).

Throughout this specification and the claims which follow, unless thecontext requires otherwise, the word “comprise”, and variations such as“comprises” and “comprising”, will be understood to imply the inclusionof a stated integer or step or group of integers or steps but not theexclusion of any other integer or step or group of integers or steps.

Several documents are cited throughout the text of this specification.Each of the documents cited herein (including all patents, patentapplications, scientific publications, manufacturer's specifications,instructions, etc.), whether supra or infra, are hereby incorporated byreference in their entirety. Nothing herein is to be construed as anadmission that the invention is not entitled to antedate such disclosureby virtue of prior invention.

In the following definitions of the terms: alkyl, heteroalkyl,heteroaryl, alkenyl, and alkynyl are provided. These terms will in eachinstance of its use in the remainder of the specification have therespectively defined meaning and preferred meanings. Nevertheless insome instances of their use throughout the specification preferredmeanings of these terms are indicated.

The term “alkyl” refers to a saturated straight or branched carbonchain. Preferably, the chain comprises from 1 to 10 carbon atoms, i.e.1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, e.g. methyl, ethyl methyl,ethyl, propyl, iso-propyl, butyl, iso-butyl, tert-butyl, pentyl, hexyl,heptyl, octyl. Alkyl groups are optionally substituted where indicated.

The term “C₁-C₄ hydroxyalkyl” refers to a mono- or poly-hydroxylatedC₁-C₄ alkyl, i.e. comprising 1, 2, 3 or 4 carbon atoms. Preferably, thisterm refers to a mono-hydroxylated C₁-C₄ alkyl group, e.g.hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyl-iso-propyl,1-hydroxybutyl or 2-hydroxybutyl.

The term “heteroalkyl” refers to a saturated straight or branched carbonchain. Preferably, the chain comprises from 1 to 9 carbon atoms, i.e. 1,2, 3, 4, 5, 6, 7, 8, 9 carbon atoms, e.g. methyl, ethyl, propyl,iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl,heptyl, octyl, which is interrupted one or more times, e.g. 1, 2, 3,with the same or different heteroatoms. Preferably the heteroatoms areselected from O, S, and N, e.g. CH₂—O—CH₃, CH₂—O—C₂H₅, C₂H₄—O—CH₃,C₂H₄—O—C₂H₅ etc. Heteroalkyl groups are optionally substituted.

The term “heteroaryl” preferably refers to a five or six-memberedaromatic monocyclic ring wherein at least one of the carbon atoms arereplaced by 1, 2, 3 or 4 (for the five membered ring) or 1, 2, 3, or 4,(for the six membered ring) of the same or different heteroatoms,preferably selected from O, N and S; Examples are furanyl, thiophenyl,oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, pyrrolyl,imidazolyl, pyrazolyl, 1,2,3-triazolyl, thiazolyl, isothiazolyl,1,2,3,-thiadiazolyl, 1,2,5-thiadiazolyl, pyridinyl, pyrimidinyl,pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl and 1,3,5-triazinyl.

As used herein, “monocyclic” refers to substituents having only onearomatic or heteroaromatic ring, excluding any multiple fused aromaticor heteroaromatic rings.

The terms “alkenyl” refer to olefinic unsaturated carbon atomscontaining one or more double bonds. One example is propenyl.Preferably, the alkenyl chain comprises from 2 to 8 carbon atoms, i.e.2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethenyl, 1-propenyl,2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, iso-butenyl,sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, hexenyl,heptenyl, octenyl.

The term “alkynyl” refers to unsaturated carbon atoms with one or moretriple bonds. An example is the propargyl radical. Preferably, thealkynyl chain comprises from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7,or 8 carbon atoms, e.g. ethynyl, 1-propynyl, 2-propynyl, 1-butynyl,2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl,hexynyl, pentynyl or octynyl.

In one preferred embodiment of the compounds of the invention, hydrogenatoms in an alkyl, aryl, alkenyl or alkynyl radical may be substitutedindependently from each other with one or more halogen atoms. Oneradical is the trifluoromethyl radical.

If two or more radicals can be selected independently from each other,then the term “independently” means that the radicals may be the same ormay be different.

The term “pharmaceutically acceptable salt” refers to a salt of thecompound of the present invention. Suitable pharmaceutically acceptablesalts of the compound of the present invention include acid additionsalts which may, for example, be formed by mixing a solution of thecompounds of the present invention with a solution of a pharmaceuticallyacceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid,maleic acid, succinic acid, acetic acid, benzoic acid, citric acid,tartaric acid, carbonic acid or phosphoric acid. Furthermore, where thecompound of the invention carries an acidic moiety, suitablepharmaceutically acceptable salts thereof may include alkali metal salts(e.g., sodium or potassium salts); alkaline earth metal salts (e.g.,calcium or magnesium salts); and salts formed with suitable organicligands (e.g., ammonium, quaternary ammonium and amine cations formedusing counteranions such as halide, hydroxide, carboxylate, sulfate,phosphate, nitrate, alkyl sulfonate and aryl sulfonate). Illustrativeexamples of pharmaceutically acceptable salts include but are notlimited to: acetate, adipate, alginate, ascorbate, aspartate,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,bromide, butyrate, calcium edetate, camphorate, camphorsulfonate,camsylate, carbonate, chloride, citrate, clavulanate,cyclopentanepropionate, digluconate, dihydrochloride, dodecylsulfate,edetate, edisylate, estolate, esylate, ethanesulfonate, formate,fumarate, gluceptate, glucoheptonate, gluconate, glutamate,glycerophosphate, glycolylarsanilate, hemisulfate, heptanoate,hexanoate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride,hydroiodide, 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide,isothionate, lactate, lactobionate, laurate, lauryl sulfate, malate,maleate, malonate, mandelate, mesylate, methanesulfonate, methylsulfate,mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate,N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate),palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate,phosphate/diphosphate, picrate, pivalate, polygalacturonate, propionate,salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate,teoclate, tosylate, triethiodide, undecanoate, valerate, and the like(see, for example, Berge, S. M., et al, “Pharmaceutical Salts”, Journalof Pharmaceutical Science, 1977, 66, 1-19). Certain specific compoundsof the present invention contain both basic and acidic functionalitiesthat allow the compounds to be converted into either base or acidaddition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide a compound of general formula(I)-(III). A prodrug is a pharmacologically active or inactive compoundthat is modified chemically through in vivo physiological action, suchas hydrolysis, metabolism and the like, into a compound of thisinvention following administration of the prodrug to a patient.Additionally, prodrugs can be converted to the compounds of the presentinvention by chemical or biochemical methods in an ex vivo environment.For example, prodrugs can be slowly converted to the compounds of thepresent invention when placed in a transdermal patch reservoir with asuitable enzyme. The suitability and techniques involved in making andusing prodrugs are well known by those skilled in the art. For a generaldiscussion of prodrugs involving esters see Svensson and Tunek DrugMetabolism Reviews 16.5 (1988) and Bundgaard Design of Prodrugs,Elsevier (1985). Examples of a masked acidic anion include a variety ofesters, such as alkyl (for example, methyl, ethyl), cycloalkyl (forexample, cyclohexyl), aralkyl (for example, benzyl, p-methoxybenzyl),and alkylcarbonyloxyalkyl (for example, pivaloyloxymethyl). Amines havebeen masked as arylcarbonyloxymethyl substituted derivatives which arecleaved by esterases in vivo releasing the free drug and formaldehyde(Bungaard J. Med. Chem. 2503 (1989)). Also, drugs containing an acidicNH group, such as imidazole, imide, indole and the like, have beenmasked with N-acyloxymethyl groups (Bundgaard Design of Prodrugs,Elsevier (1985)). Hydroxy groups have been masked as esters and ethers.EP 0 039 051 (Sloan and Little, Apr. 11, 1981) discloses Mannich-basehydroxamic acid prodrugs, their preparation and use.

Compounds of the present invention and also the starting materials fortheir preparation according to the invention can be synthesized as shownherein, and, alternatvely, by methods and standard procedures known tothose skilled in the art, i.e. as described in the literature (forexample in the standard works, such as Houben-Weyl, Methoden derorganischen Chemie [Methods of Organic Chemistry], Georg-Thieme-Verlag,Stuttgart), under reaction conditions which are known to those skilledin the art and suitable for the said reactions.

If desired, the starting materials can also be formed in situ by notisolating them from the reaction mixture, but instead immediatelyconverting them further into the compounds of the present invention. Onthe other hand, it is possible to carry out the reaction stepwise. Itshould be noted that the general procedures are shown as it relates topreparation of compounds having unspecified stereochemistry. However,such procedures are generally applicable to those compounds of aspecific stereochemistry, e.g., where the stereochemistry at asterogenic center is (S) or (R). In addition, the compounds having onestereochemistry (e.g., (R)) can often be utilized to produce thosehaving opposite stereochemistry (i.e., (S)) using well-known methods,for example, by inversion.

Certain compounds of the present invention can exist in unsolvated formsas well as in solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds. The racemates, enantiomers,diastereomers, geometric isomers and individual isomers are all intendedto be encompassed within the scope of the present invention.Accordingly, the compounds of this invention include mixtures ofstereoisomers, especially mixtures of enantiomers, as well as purifiedstereoisomers, especially purified enantiomers, or stereoisomericallyenriched mixtures, especially enantiomerically enriched mixtures. Alsoincluded within the scope of the invention are the individual isomers ofthe compounds represented by formulas (I) to (III) below as well as anywholly or partially equilibrated mixtures thereof. The present inventionalso covers the individual isomers of the compounds represented by theformulas below as mixtures with isomers thereof in which one or morechiral centers are inverted. Also, it is understood that all tautomersand mixtures of tautomers of the compounds of formulas (I) to (III) areincluded within the scope of the compounds of formulas (I) to (III) andpreferably the formulas and subformulas corresponding thereto.

Racemates obtained can be resolved into the isomers mechanically orchemically by methods known per se. Diastereomers are preferably formedfrom the racemic mixture by reaction with an optically active resolvingagent.

Examples of suitable resolving agents are optically active acids, suchas the D and L forms of tartaric acid, diacetyltartaric acid,dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or thevarious optically active camphorsulfonic acids, such as—camphorsulfonicacid. Also advantageous is enantiomer resolution with the aid of acolumn filled with an optically active resolving agent (for exampledinitrobenzoylphenylglycine); an example of a suitable eluent is ahexane/isopropanol/acetonitrile mixture.

The diastereomer resolution can also be carried out by standardpurification processes, such as, for example, chromatography orfractional crystallization.

It is also possible to obtain optically active compounds of formulas (I)to (III) by the methods described above by using starting materialswhich are already optically active.

The present invention provides novel compositions, compounds and uses ofthese compounds and compositions for the prevention or treatment of adisease or disorder.

In a first aspect the present invention relates to a compound having astructure according to formula I:

wherein

-   R¹ is selected from a group consisting of H; alkyl, preferably the    alkyl chain comprises from 1 to 10 carbon atoms, i.e. 1, 2, 3, 4, 5,    6, 7, 8, 9 or 10 carbon atoms e.g. methyl, ethyl, propyl,    iso-propyl, butyl, iso-butyl, tert-butyl, pentyl, hexyl, heptyl or    octyl; alkenyl, particularly an alkenyl chain comprising from 2 to 8    carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g.    ethenyl, 1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl,    3-butenyl, iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl,    3-pentenyl, 4-pentenyl, hexenyl, heptenyl or octenyl; alkynyl,    preferably an alkynyl comprising from 2 to 8 carbon atoms, i.e. 2,    3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethynyl, 1-propynyl,    2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl,    3-pentynyl, 4-pentynyl, hexynyl, pentynyl or octynyl; —CN; halogen,    in particular F, Cl, Br or I; —OH; alkoxy, in particular C₁-C₆    alkoxy, e.g. C₁, C₂, C₃, C₄, C₅, or C₆ alkoxy, preferably methoxy,    ethoxy, propoxy, iso-propoxy, butoxy, iso-butoxy, tert-butoxy,    pentoxy, or hexoxy; —SH; S-alkyl, in particular C₁-C₆ S-alkyl, e.g.    C_(I), C₂, C₃, C₄, C₅, or C₆ S-alkyl; —NH₂; NH-alkyl, in particular    C₁-C₆ NH-alkyl, e.g. C₁, C₂, C₃, C₄, C₅, or C₆ NH-alkyl;    N-bis-alkyl, preferably C₁-C₆ N-bis-alkyl, e.g. C₁, C₂, C₃, C₄, C₅,    or C₆ N-bis-alkyl; NHOH; NMeOH; NMe(OMe); —NO₂; —CF₃; —OCF₃ and    C₁-C₄ hydroxyalkyl, in particular C₁-C₄ hydroxyalkyl, e.g. C₁, C₂,    C₃ or C₄ hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,    hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or    2-hydroxybutyl;-   R² is H or C₁-C₄ alkyl, i.e. an alkyl group comprising from 1 to 4    carbon atoms, i.e. 1, 2, 3 or 4 carbon atoms e.g. methyl, ethyl,    propyl, iso-propyl, butyl or iso-butyl;-   R³ is H or —CH₃;-   R⁴ is phenyl or monocyclic 5- or 6-membered heteroaryl; optionally    substituted with one or more substituents selected from the group    consisting of:    -   alkyl, preferably the alkyl the chain comprises from 1 to 10        carbon atoms, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms        e.g. methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl,        tert-butyl, pentyl, hexyl, heptyl or octyl; alkenyl,        particularly an alkenyl chain comprising from 2 to 8 carbon        atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethenyl,        1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl,        3-butenyl, iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl,        3-pentenyl, 4-pentenyl, hexenyl, heptenyl or octenyl; alkynyl,        preferably an alkynyl comprising from 2 to 8 carbon atoms, i.e.        2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethynyl, 1-propynyl,        2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl,        2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, pentynyl or        octynyl; alkoxy, in particular C₁-C₆ alkoxy, e.g. C₁, C₂, C₃,        C₄, C₅, or C₆ alkoxy, preferably methoxy, ethoxy, propoxy,        iso-propoxy, butoxy, iso-butoxy, tert-butoxy, pentoxy, or        hexoxy; halogen, in particular F, Cl, Br or I; —CN; —CF₃; —OCF₃;        C₁-C₄ hydroxyalkyl, in particular C₁-C₄ hydroxyalkyl, e.g. C₁,        C₂, C₃ or C₄ hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,        hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or        2-hydroxybutyl; —OH; —SH; S-alkyl, in particular C₁-C₆ S-alkyl,        e.g. C₁, C₂, C₃, C₄, C₅, or C₆ S-alkyl; —CN; N-bis-alkyl,        preferably C₁-C₆N-bis-alkyl, e.g. C₁, C₂, C₃, C₄, C₅, or C₆        N-bis-alkyl; cyanoacetylene; —NO₂; —NR⁷R⁸, preferably —NH₂ or        NH-alkyl, more preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃,        —NH-ethyl, —NH—C₃ alkyl or —NH—C₄ alkyl and most preferably        —NH₂, NH(CH₃) or N(CH₃)₂; —C(O)R²⁰, preferably acetyl,        propionyl, iso-propionyl, butyryl or iso-butyryl; N-0 (wherein        the nitrogen atom is integral part of the monocyclic 5- or        6-membered heteroaryl) and two substituents which form together        a dioxymethylene bridge (—O—CH₂—O—);-   R⁵ is H or —CH₃;-   R⁶ is selected from the group consisting of H; halogen, in    particular F, Cl, Br or I; alkyl, preferably the alkyl the chain    comprises from 1 to 10 carbon atoms, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9    or 10 carbon atoms e.g. methyl, ethyl, propyl, iso-propyl, butyl,    iso-butyl, tert-butyl, pentyl, hexyl, heptyl or octyl; alkoxy, in    particular C₁-C₆ alkoxy, e.g. C₁, C₂, C₃, C₄, C₅, or C₆ alkoxy,    preferably methoxy, ethoxy, propoxy, iso-propoxy, butoxy,    iso-butoxy, tert-butoxy, pentoxy, or hexoxy; alkenyl, particularly    an alkenyl chain comprising from 2 to 8 carbon atoms, i.e. 2, 3, 4,    5, 6, 7, or 8 carbon atoms, e.g. ethenyl, 1-propenyl, 2-propenyl,    iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, iso-butenyl,    sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl,    hexenyl, heptenyl or octenyl; alkynyl, preferably an alkynyl    comprising from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8    carbon atoms, e.g. ethynyl, 1-propynyl, 2-propynyl, 1-butynyl,    2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl,    4-pentynyl, hexynyl, pentynyl or octynyl; S-alkyl, in particular    C₁-C₆ S-alkyl, e.g. C₁, C₂, C₃, C₄, C₅, or C₆ S-alkyl; —OH; —NR⁷R⁸,    preferably —NH₂ or NH-alkyl, more preferably —NH₂ or —NH—C₁-C₄    alkyl, i.e. —NH—CH₃, —NH-ethyl, —NH—C₃ alkyl or —NH—C₄ alkyl and    most preferably —NH₂, NH(CH₃) or N(CH₃)₂; —CN; N-bis-alkyl,    preferably C₁-C₆ N-bis-alkyl, e.g. C₁, C₂, C₃, C₄, C₅, or C₆    N-bis-alkyl; —SH; —CF₃ and —OCF₃;    -   or R⁶ forms together with R¹ a dioxymethylene bridge        (—O—CH₂—O—), optionally substituted at the methylene;-   R⁷ is H or alkyl, preferably the alkyl the chain comprises from 1 to    10 carbon atoms, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms    e.g. methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl,    tert-butyl, pentyl, hexyl, heptyl or octyl;-   R⁸ is H or C₁-C₄ alkyl, in particular methyl, ethyl, propyl,    iso-propyl, butyl or iso-butyl; and-   R²⁰ is C₁-C₄ alkyl, in particular methyl, ethyl, propyl, iso-propyl,    butyl or iso-butyl;    with the proviso that R⁴ is not 3-alkoxy-pyridazine-5-yl; if R⁴ is    phenyl, then the 2- and 5-position of the phenyl ring may not be    substituted with two methoxy substituents at the same time; and that    R³ and R⁵ are not at the same time H.

Also preferred is a compound, wherein R¹ is selected from the groupconsisting of —CH₃, —OCH₃, —CF₃, F and —NH₂.

In a further preferred embodiment of the compound, R⁶, R² and R⁵ is Hand R¹ is selected from the group consisting of —CH₃, —OCH₃, —CF₃, F and—NH₂.

The R⁴ substituent of a compound of the invention may have in preferredembodiments only a single heteroatom and/or may be substituted with lessthan two or less than three substituents.

A further preferred embodiment of the compound of the invention has thefollowing substituents: R⁶, R² and R⁵ is H and R¹ is selected from thegroup consisting of —CH₃, —OCH₃, —CF₃, F and —NH₂ and the R⁴ group issubstituted with less than two or less than three substituents and/or,if R⁴ is a monocyclic 5- or 6-membered heteroaryl, then it is preferredin this context that it comprises only a single heteroatom.

In a preferred embodiment the compound of the invention has a structureaccording to formula II:

Also preferred is a compound of the invention having a structureaccording to formula III:

In the context of formulas (II) and (III) R¹, R², R⁴ and R⁶ have theabove indicated meaning and preferred meanings. Particularly preferredis a compound of formula (II) or (III), wherein R¹ is selected from thegroup consisting of —CH₃, —OCH₃, —CF₃, F and —NH₂. In a furtherpreferred embodiment of the compound according to formula (II) or (III)R² is H. In an even more preferred embodiment R⁶ and R² are H and R¹ isselected from the group consisting of —CH₃, —OCH₃, —CF₃, F and —NH₂.

In a preferred embodiment of the compound of the invention, preferably acompound according to formula (II) or (III), R⁴ has a structureaccording to formula IV:

wherein

-   R⁹ and R¹⁰ are each individually selected from the group consisting    of H; C₁-C₄ alkyl, in particular methyl, ethyl, propyl, iso-propyl,    butyl or iso-butyl; C₁-C₄ alkenyl, e.g. ethenyl, 1-propenyl,    2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,    iso-butenyl or sec-butenyl; C₁-C₄ alkynyl, e.g. ethynyl, 1-propynyl,    2-propynyl, 1-butynyl, 2-butynyl or 3-butynyl; —CN; —C(O)R²⁰,    preferably acetyl, propionyl, iso-propionyl, butyryl or iso-butyryl;    cyanoacetylene; halogen, in particular F, Cl, Br or I; —OH; C₁-C₄    alkoxy, e.g. methoxy, ethoxy, propoxy, iso-propoxy, butoxy,    iso-butoxy or tert-butoxy; —SH; C₁-C₄ S-alkyl, e.g. C₁, C₂, C₃ or C₄    S-alkyl; —NH₂; C₁-C₄ NH-alkyl, e.g. C₁, C₂, C₃ or C₄ NH-alkyl; C₁-C₄    N-bis-alkyl, e.g. C₁, C₂, C₃ or C₄ N-bis-alkyl; —NO₂; —CF₃; —OCF₃;    and C₁-C₄ hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,    hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or    2-hydroxybutyl; or R⁹ and R¹⁰ form together a dioxymethylene bridge    (—O—CH₂—O—);-   R¹¹ and R¹² are each individually selected from the group consisting    of H; C₁-C₄ alkyl in particular methyl, ethyl, propyl, iso-propyl,    butyl or iso-butyl; C₁-C₄ alkenyl, e.g. ethenyl, 1-propenyl,    2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,    iso-butenyl or sec-butenyl; C₁-C₄ alkynyl, e.g. ethynyl, 1-propynyl,    2-propynyl, 1-butynyl, 2-butynyl or 3-butynyl; —CN; halogen, in    particular F, Cl, Br or I; —OH; C₁-C₄ alkoxy, e.g. methoxy, ethoxy,    propoxy, iso-propoxy, butoxy, iso-butoxy or tert-butoxy; —SH; C₁-C₄    S-alkyl, e.g. C₁, C₂, C₃ or C₄ S-alkyl; —CF₃; —OCF₃; —NH₂; —N(CH₃)₂    and C₁-C₄ hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,    hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or    2-hydroxybutyl;    with the proviso that R⁹ and R¹² may not be methoxy at the same    time;-   R²⁰ has the above indicated meaning, e.g. methyl, ethyl, propyl,    iso-propyl, butyl or iso-butyl; and-   * indicates the bond between R⁴ and the compound according to any of    formulas (I) through (III).

In the preferred embodiment wherein R⁴ has the structure of formula (IV)it is particularly preferred that (a) R⁹ is CN, R¹⁰, R¹¹ and R¹² is H;(b) R¹⁰ is CN and R⁹, R¹¹ and R¹² is H, (c) R⁹, R¹⁰ and R¹¹ is H and R¹²is CN; (d) R⁹ is ethynyl, R¹⁰, R¹¹ and R¹² is H; (e) R¹⁰ is ethynyl, R⁹,R¹¹ and R¹² is H, (f) R⁹, R¹⁰ and R¹¹ is H and R¹² is ethynyl; (g) R⁹ isCN and one of R¹⁰, R¹¹ and R¹² is OH or C₁-C₄ alkoxy, e.g. methoxy,ethoxy, propoxy, iso-propoxy, butoxy, iso-butoxy or tert-butoxy; (h) R¹⁰is CN and one of R⁹, R¹¹ and R¹², preferably R⁹ is halogen, inparticular F, OH or C₁-C₄ alkoxy, e.g. methoxy, ethoxy, propoxy,iso-propoxy, butoxy, iso-butoxy or tert-butoxy or (i) one of R⁹, R¹⁰ andR¹¹ is OH or C₁-C₄ alkoxy, e.g. methoxy, ethoxy, propoxy, iso-propoxy,butoxy, iso-butoxy or tert-butoxy and R¹² is CN; preferably R¹⁰ is CNand R⁹ is OH or C₁-C₄ alkoxy, preferably methoxy.

Further preferred is a compound of the invention, wherein R¹¹ and R¹²are H.

In another preferred embodiment of the compound of the invention, R³ ismethyl; R² is H, methyl or ethyl, preferably H; and R⁵ and R⁶ are H.

In a preferred embodiment of the compound of the invention, R⁴ has astructure according to formula V:

wherein

-   A, B, D and E are each individually selected from the group    consisting of a nitrogen atom, CR¹³ and N—O;-   G is selected from the group consisting of an oxygen atom, a sulphur    atom and NR¹⁴;-   R¹³ is selected from the group consisting of H; C₁-C₃ alkyl, in    particular methyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, e.g.    methoxy, ethoxy, propoxy or iso-propoxy; —OH; —SH; S-alkyl, e.g. C₁,    C₂ or C₃ S-alkyl; —CF₃; —OCF₃; halogen, in particular F, Cl, Br or    I; —NR¹⁵R¹⁶, preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃,    —NH-ethyl, —NH—C₃ alkyl or —NH—C₄ alkyl and most preferably —NH₂,    NH(CH₃) or N(CH₃)₂; —NO₂; —CN; —C(O)R²⁰; acetylene; cyanoacetylene;    C₁-C₄ hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl, hydroxypropyl    or hydroxyl-iso-propyl, 1-hydroxybutyl or 2-hydroxybutyl and a σ    (sigma) bond connecting R⁴ to the compound according to any of    formulas (I) through (III); and-   R¹⁴ is selected from the group consisting of H, C₁-C₄ alkyl, in    particular methyl, ethyl, propyl, iso-propyl, butyl or iso-butyl and    more preferably methyl, ethyl, propyl or iso-propyl; and a σ (sigma)    bond connecting R⁴ to the compound according to any of formulas (I)    through (III); and-   R¹⁵ and R¹⁶ are each individually either H or C₁-C₄ alkyl, in    particular methyl, ethyl, propyl, iso-propyl, butyl or iso-butyl;-   R²⁰ has the above indicated meaning, e.g. methyl, ethyl, propyl,    iso-propyl, butyl or iso-butyl; and-   * indicates the bond between R⁴ and the compound according to any of    formulas (I) through (III).

In another preferred embodiment of the compound of the invention, R⁴ hasa structure according to formula VI:

wherein

-   L and T are each individually either a CH group or a nitrogen atom    or N—O;-   M, N and Q are each individually selected from the group consisting    of a nitrogen atom, a CR¹⁷ group and N—O;-   R¹⁷ is selected from the group consisting of H; C₁-C₃ alkyl, in    particular methyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, e.g.    methoxy, ethoxy, propoxy or iso-propoxy; —CF₃; —OCF₃; halogen, in    particular F, Cl, Br or I; —OH; —NO₂; —SH; C₁-C₃ S-alkyl, e.g. C₁,    C₂ or C₃ S-alkyl; —NR¹⁵R¹⁶, preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e.    —NH—CH₃, —NH-ethyl, —NH—C₃ alkyl or —NH—C₄ alkyl and most preferably    —NH₂, NH(CH₃) or N(CH₃)₂; C₁-C₄ hydroxyalkyl, e.g. hydroxymethyl,    hydroxyethyl, hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl    or 2-hydroxybutyl; —C(O)R²⁰, preferably acetyl, propionyl,    iso-propionyl, butyryl or iso-butyryl; acetylene; cyanoacetylene and    —CN;-   R¹⁵ and R¹⁶ are each individually either H or C₁-C₄ alkyl;-   R²⁰ has the above indicated meaning, e.g. methyl, ethyl, propyl,    iso-propyl, butyl or iso-butyl; and-   * indicates the bond between R⁴ and the compound according to any of    formulas (I) through (III).

Also preferred is a compound of the invention, wherein R⁴ is selectedfrom the group consisting of:

-   wherein R¹⁸ and R¹⁹ are each individually selected from the group    consisting of H; C₁-C₃ alkyl, in particular methyl, ethyl, propyl or    iso-propyl; C₁-C₃ alkoxy, e.g. methoxy, ethoxy, propoxy or    iso-propoxy; —CF₃; —OCF₃; halogen, in particular F, Cl, Br or I;    —OH; —NO₂; —SH; C₁-C₃ S-alkyl, e.g. C₁, C₂ or C₃ S-alkyl; —NR¹⁵R¹⁶,    preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃, —NH-ethyl,    —NH—C₃-alkyl or —NH—C₄-alkyl and most preferably —NH₂, NH(CH₃) or    N(CH₃)₂; C₁-C₄-hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,    hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or    2-hydroxybutyl; alkenyl, particularly an alkenyl chain comprising    from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms,    e.g. ethenyl, 1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl,    2-butenyl, 3-butenyl, iso-butenyl, sec-butenyl, 1-pentenyl,    2-pentenyl, 3-pentenyl, 4-pentenyl, hexenyl, heptenyl or octenyl;    alkynyl, preferably an alkynyl comprising from 2 to 8 carbon atoms,    i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethynyl, 1-propynyl,    2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl,    3-pentynyl, 4-pentynyl, hexynyl, pentynyl or octynyl; —C(O)R⁶    preferably acetyl, propionyl, iso-propionyl, butyryl or iso-butyryl;    cyanoacetylene and —CN;-   and R¹⁵ and R¹⁶ have the above indicated meaning and preferred    meaning.

In a preferred embodiment R¹⁸ is H and R¹⁹ in above indicated structuresis selected from the group consisting of C₁-C₃ alkyl, in particularmethyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, e.g. methoxy, ethoxy,propoxy or iso-propoxy; —CF₃; —OCF₃; halogen, in particular F, Cl, Br orI; —OH; —NO₂; —SH; C₁-C₃ S-alkyl, e.g. C₁, C₂ or C₃ S-alkyl; —NR¹⁵R¹⁶,preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃, —NH-ethyl,—NH—C₃-alkyl or —NH—C₄-alkyl and most preferably —NH₂, NH(CH₃) orN(CH₃)₂; C₁-C₄-hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or 2-hydroxybutyl;alkenyl, particularly an alkenyl chain comprising from 2 to 8 carbonatoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethenyl,1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,4-pentenyl, hexenyl, heptenyl or octenyl; alkynyl, preferably an alkynylcomprising from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbonatoms, e.g. ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl,3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl,pentynyl or octynyl; —C(O)R⁶ preferably acetyl, propionyl,iso-propionyl, butyryl or iso-butyryl; cyanoacetylene and —CN. The mostpreferred meanings of R¹⁹ in this context are C₁-C₃ alkyl, in particularmethyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, in particularmethoxy, ethoxy, propoxy or iso-propoxy; —CF₃; —OCF₃, —CN, —NO₂, orhalogen, in particular F, Cl, Br or I, with methyl, methoxy, CF₃ or —CNbeing the most preferred meanings.

In another preferred embodiment R¹⁹ is H and R¹⁸ in above indicatedstructures is selected from the group consisting of C₁-C₃ alkyl, inparticular methyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, e.g.methoxy, ethoxy, propoxy or iso-propoxy; —CF₃; —OCF₃; halogen, inparticular F, Cl, Br or I; —OH; —NO₂; —SH; C₁-C₃ S-alkyl, e.g. C₁, C₂ orC₃ S-alkyl; —NR¹⁵R¹⁶, preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃,—NH-ethyl, —NH—C₃-alkyl or —NH—C₄-alkyl and most preferably —NH₂,NH(CH₃) or N(CH₃)₂; C₁-C₄-hydroxyalkyl, e.g. hydroxymethyl,hydroxyethyl, hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or2-hydroxybutyl; alkenyl, particularly an alkenyl chain comprising from 2to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g.ethenyl, 1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl,3-butenyl, iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,4-pentenyl, hexenyl, heptenyl or octenyl; alkynyl, preferably an alkynylcomprising from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbonatoms, e.g. ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl,3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl,pentynyl or octynyl; —C(O)R⁶ preferably acetyl, propionyl,iso-propionyl, butyryl or iso-butyryl; cyanoacetylene and —CN. The mostpreferred meanings of R¹⁸ in this context are C₁-C₃ alkyl, in particularmethyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, in particularmethoxy, ethoxy, propoxy or iso-propoxy; —CF₃; —OCF₃, —CN, NO₂, orhalogen, in particular F, Cl, Br or I, with methyl, methoxy, CF₃ or —CNbeing the most preferred meanings.

In another preferred embodiment R¹⁸ and R¹⁹ are each individuallyselected from the group consisting of C₁-C₃ alkyl, in particular methyl,ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, e.g. methoxy, ethoxy, propoxyor iso-propoxy; —CF₃; —OCF₃; halogen, in particular F, Cl, Br or I; —OH;—NO₂; —SH; C₁-C₃ S-alkyl, e.g. C₁, C₂ or C₃ S-alkyl; —NR¹⁵R¹⁶,preferably —NH₂ or —NH—C₁-C₄ alkyl, i.e. —NH—CH₃, —NH-ethyl,—NH—C₃-alkyl or —NH—C₄-alkyl and most preferably —NH₂, NH(CH₃) orN(CH₃)₂; C₁-C₄-hydroxyalkyl, e.g. hydroxymethyl, hydroxyethyl,hydroxypropyl or hydroxyl-iso-propyl, 1-hydroxybutyl or 2-hydroxybutyl;alkenyl, particularly an alkenyl chain comprising from 2 to 8 carbonatoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbon atoms, e.g. ethenyl,1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl,iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,4-pentenyl, hexenyl, heptenyl or octenyl; alkynyl, preferably an alkynylcomprising from 2 to 8 carbon atoms, i.e. 2, 3, 4, 5, 6, 7, or 8 carbonatoms, e.g. ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl,3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl,pentynyl or octynyl; —C(O)R⁶ preferably acetyl, propionyl,iso-propionyl, butyryl or iso-butyryl; cyanoacetylene and —CN. The mostpreferred meanings of R¹⁸ and R¹⁹ in this context are C₁-C₃ alkyl, inparticular methyl, ethyl, propyl or iso-propyl; C₁-C₃ alkoxy, inparticular methoxy, ethoxy, propoxy or iso-propoxy; —CF₃; —OCF₃, —CN,NO₂, or halogen, in particular F, Cl, Br or I, with methyl, methoxy, CF₃or —CN being the most preferred meanings.

In another preferred embodiment of the compound of the invention, R² isH and R⁴ is selected from the group consisting of:

wherein * indicates the bond between R⁴ and the compound according toany of formulas (I) through (III).

In a further preferred embodiment of the compound of the invention R²and/or R⁶ is H.

The following Table 1 lists further preferred combinations of specificsubstituents of the compound of the invention:

TABLE 1 R¹ R² R³ R⁴ R⁵ R⁶ R¹¹ R¹² as defined H methyl as defined H H asas herein herein defined defined herein herein as defined H methyl asdefined H as defined as as herein herein herein defined defined hereinherein as defined H as defined as defined as defined H as as hereinherein herein herein defined defined herein herein as defined H asdefined as defined as defined as defined as as herein herein hereinherein herein defined defined herein herein as defined as defined asdefined as defined as defined H as as herein herein herein herein hereindefined defined herein herein as defined H methyl as defined H H H Hherein herein as defined H methyl as defined H as defined H H hereinherein herein as defined H as defined as defined as defined H H H hereinherein herein herein as defined H as defined as defined as defined asdefined H H herein herein herein herein herein as defined as defined asdefined as defined as defined H H H herein herein herein herein herein

In a particularly preferred embodiment of the compound of the invention,the compound is selected from the group of compounds listed in Table 2Aor Table 7.

In a further aspect the invention provides a compound according to theinvention or pharmaceutically acceptable salt thereof, for theprevention or treatment of a disease or disorder.

As shown in the examples below, the advantageous properties of thecompounds of the invention include their ability of effectivelyinhibiting cell proliferation and their activity as HIF inhibitors. Forexample, the compounds of the present invention were shown to inhibitthe activation of HIF-mediated transcription under hypoxic conditions.Thus, the compounds of the invention can be used for the preparation ofa medicament for the treatment of a disorder characterized bypathophysiological HIF signaling. A person skilled in the art ofmedical, biological and/or pharmacological science can determine withroutine methodology if a disorder is characterized by undesirable HIFsignaling. Tissues affected by such diseases will overexpress genes thatare induced by activation of the HIF responsive element (HRE). HIF-1acts by binding to HIF-responsive elements (HREs) in promoters thatgenerally contain the sequence NCGTG. The genes affected by HIF activitywhich are regulated by said promoters are well known in the art and werealso described in multiple reviews (see e.g. FIG. 3 of Gregg L. Semenza,Nature Reviews, October 2003, vol. 3).

In animal studies, HIF-1 overexpression is associated with increasedtumor growth, increased vascularisation, metastasis and fibrosis, e.g.renal fibrosis (see: Semenza, G, Drug Discovery Today, vol. 12, no.19/20, October 2007; Kimura, Kuniko, et al., American Journal ofPhysiology (2008), 295(4, Pt. 2), F1023-F1029 and for a review see N. J.Mabjeesh et al., Histol. Histopathol (2007) 22:559-572). Fibrosis is theformation or development of excess fibrous connective tissue in an organor tissue. Recently, it has become clear that inhibition of HIF-1activity also acts to prevent inflammation, by virtue of its essentialrole in the activation and infiltration of macrophages and neutrophilsinto affected tissues (see e.g. Giaccia et al., Drug Discovery, vol. 2,October 2003).

Pharmaceutical Compositions

For the above mentioned reasons, a compound of the present invention canbe used to treat an inflammatory disease, a hyperproliferative diseaseor disorder, a hypoxia related pathology and also diseases characterizedby pathophysiological hyper-vascularisation. Therefore, as a furtheraspect, the invention provides a therapeutical composition wherein thecompound of the invention is combined with at least one furtherpharmaceutically active compound that is useful to treat one of theaforementioned diseases or disorders. Such therapeutical compositionsare useful because the therapeutic efficiency of the compounds of theinvention can be amplified by the presence of said at least one furtherpharmaceutically active compound and vice versa. For example, it wasshown that inhibiting HIF1α activity via antisense gene therapy enhancesthe therapeutic efficacy of doxorubicin to combat hepatocellularcarcinoma (see Liu, Fengjun et. al., Cancer Science (2008), 99(10),2055-2061).

Thus, in a further aspect, the present invention relates to apharmaceutical composition comprising a compound according to theinvention or pharmaceutically acceptable salt thereof and a secondtherapeutic agent useful for the treatment or prevention of a disease ordisorder selected from the group consisting of an inflammatory disease,a hyperproliferative disease or disorder, a hypoxia related pathologyand a disease characterized by pathophysiological hyper-vascularisation,and, optionally, a pharmaceutically acceptable carrier or excipient.Such compositions are also useful to obtain synergistic therapeuticeffects and also to prevent drug resistance of tumor cells, for example.It is also for these reasons, that current chemotherapy generallyinvolves administering a cocktail of different cytotoxic and/orcytostatic compounds to improve the effectiveness of the treatment andreduce the possibility of tumor cell adaptation. In a further aspect,the present invention relates to a pharmaceutical composition comprisinga compound according to the invention or pharmaceutically acceptablesalt thereof in combination with radiation therapies.

Any composition of the present invention may be admixed with apharmaceutically acceptable diluent, excipient or carrier, or a mixturethereof.

Even though the compounds of the present invention (including theirpharmaceutically acceptable salts, esters and pharmaceuticallyacceptable solvates) can be administered alone, they will generally beadministered in admixture with a pharmaceutical carrier, excipient ordiluent, particularly for human therapy. The pharmaceutical compositionsmay be for human or animal usage in human and veterinary medicine.Examples of such suitable excipients for the various different forms ofpharmaceutical compositions described herein may be found in the“Handbook of Pharmaceutical Excipients”, 2nd Edition, (1994), Edited byA Wade and P J Weller. Acceptable carriers or diluents for therapeuticuse are well known in the pharmaceutical art, and are described, forexample, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A.R Gennaro edit. 1985).

For preparing pharmaceutical compositions from the compounds of thepresent invention, pharmaceutically acceptable carriers can be eithersolid or liquid. Solid form preparations include powders, tablets,pills, capsules, cachets, suppositories, and dispersible granules. Asolid carrier can be one or more substances, which may also act asdiluents, flavoring agents, binders, preservatives, tabletdisintegrating agents, or an encapsulating material.

In powders, the carrier is a finely divided solid, which is in a mixturewith the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary bindingproperties in suitable proportions and compacted in the shape and sizedesired.

The powders and tablets preferably contain from 5% to 80%, morepreferably from 20% to 70% of the active compound or active compounds.Suitable carriers are magnesium carbonate, magnesium stearate, talc,sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth,methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoabutter, and the like. The term “preparation” is intended to include theformulation of the active compound with encapsulating material as acarrier providing a capsule in which the active component with orwithout other carriers, is surrounded by a carrier, which is thus inassociation with it. Similarly, cachets and lozenges are included.Tablets, powders, capsules, pills, cachets, and lozenges can be used assolid dosage forms suitable for oral administration.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool, and thereby to solidify.

Liquid form preparations include solutions, suspensions, and emulsions,for example, water or water/propylene glycol solutions. Liquid forms areparticularly preferred for topical applications to the eye. Forparenteral injection, liquid preparations can be formulated in solutionin aqueous polyethylene glycol solution.

Aqueous solutions suitable for oral use can be prepared by dissolvingthe active component in water and adding suitable colorants, flavors,stabilizers, and thickening agents as desired. Aqueous suspensionssuitable for oral use can be made by dispersing the finely dividedactive component in water with viscous material, such as natural orsynthetic gums, resins, methylcellulose, sodium carboxymethylcellulose,and other well-known suspending agents.

Also included are solid form preparations, which are intended to beconverted, shortly before use, to liquid form preparations for oraladministration. Such liquid forms include solutions, suspensions, andemulsions. These preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

The pharmaceutical preparation is preferably in unit dosage form. Insuch form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Interestingly, HIF inhibitors, such as the compounds of the invention,can prevent the development of tumor resistance towards chemotherapeuticdrugs and can make cancer cells more sensitive towards radiotherapy (seee.g. Palayoor S T, et al., Int J Cancer. 2008 Nov. 15; 123(10):2430-7and Gregg L. Semenza, Nature Reviews, October 2003, vol. 3). Thus,useful second therapeutic agents that can be combined with a compound ofthe invention to produce the pharmaceutical composition of the inventioninclude, without limitation, a (further) HIF-1 inhibitor, a cytotoxiccompound and cytostatic compounds.

A HIF-1 inhibitor can be, e.g. selected from the group consisting ofPX-478 (S-2-amino-3-[4′-N,N,-bis(2-chloroethyl)amino]phenyl propionicacid N-oxide dihydrochloride); a topoisomerase-1 inhibitor such as8,9-Dimethoxy-5-(2-N,N-dimethylaminoethyl)-2,3-methylenedioxy-5H-dibenzo[c,h][1,6]naphthyridin-6-one(also known as ARC-111 or topovale) or(S)-10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3′,4′:6,7]indolizino[1,2-1)]quinoline-3,14-(4H, 12H)-dione monohydrochloride (also referred toas tropotecan); echinomycin; chetomin (NSC289491); cyclosporine A;3-[2-[4-[bis(4-fluorophenyl)methylene]-1-piperidinyl]-2,3-dihydro-2thioxo-4(1H)-quinazolinone(R59949); an inhibitor of the PIK3K/Akt/mTor signalling cascade, e.g.,LY294002, wortmannin or rapamycin; an inhibitor of the MAPK signallingcascade, e.g. the MEK1 inhibitor PD98059; a soluble guanyl cyclasestimulator such as 3-(5′ hydroxymethyl-2′-furyl)-1-benzylindazole(YC-1); a heat-shock protein 90 inhibitor, in particular radicicol, theradicicol analogue KF58333 or geldanamycin; a microtubule disruptingagent, in particular e.g. taxol, vincristine or 2-methoxyestradiol; ahistone deacetylase inhibitor, e.g. FK228; a thioredoxin inhibitor, inparticular PX-12 or pleurotin; UCNO-1; diphenylene iodonium, genesteinand carboxyamido-triazole.

Many cytotoxic or cytostatic compounds are known to the expert artisanskilled in the therapy of hyperproliferative diseases or disorders suchas a tumor or cancer disease. For example, cytotoxic and cytostaticcompounds include, but are not limited to, pure or mixed anti-estrogenssuch as faslodex, tamoxifen or raloxifen; any inhibitors oftopoisomerase I or II, such as camptothecin (topo I) or etoposide (topoII); any compound that acts through inhibiting aromatase activity, suchas anastrozole or letrozole; any preparation that interferes with HER2signalling such as herceptin; any compound that interchelates DNA, suchas doxorubicin. Particularly preferred cytostatic or cytotoxic drugs,which can be combined with the compounds of the present invention arealkylating substances, anti-metabolites, antibiotics, epothilones,nuclear receptor agonists and antagonists, anti-androgenes,anti-estrogens, platinum compounds, hormones and antihormones,interferons and inhibitors of cell cycle-dependent protein kinases(CDKs), inhibitors of cyclooxygenases and/or lipoxygenases, biogeneicfatty acids and fatty acid derivatives, including prostanoids andleukotrienes, inhibitors of protein kinases, inhibitors of proteinphosphatases, inhibitors of lipid kinases, platinum coordinationcomplexes, ethyleneimenes, methylmelamines, trazines, vinca alkaloids,pyrimidine analogs, purine analogs, alkylsulfonates, folic acid analogs,anthracendiones, substituted urea, methylhydrazin derivatives, inparticular acediasulfone, aclarubicine, ambazone, aminoglutethimide,L-asparaginase, azathioprine, bleomycin, busulfan, calcium folinate,carboplatin, carpecitabine, carmustine, celecoxib, chlorambucil,cis-platin, cladribine, cyclophosphamide, cytarabine, dacarbazine,dactinomycin dapsone, daunorubicin, dibrompropamidine,diethylstilbestrole, docetaxel, doxorubicin, enediynes, epirubicin,epothilone B, epothilone D, estramucin phosphate, estrogen,ethinylestradiole, etoposide, flavopiridol, floxuridine, fludarabine,fluorouracil, fluoxymesterone, flutamide fosfestrol, furazolidone,gemcitabine, gonadotropin releasing hormone analog, hexamethylmelamine,hydroxycarbamide, hydroxymethylnitrofurantoin,hydroxyprogesteronecaproat, hydroxyurea, idarubicin, idoxuridine,ifosfamide, interferon γ, irinotecan, leuprolide, lomustine, lurtotecan,mafenide sulfate olamide, mechlorethamine, medroxyprogesterone acetate,megastrolacetate, melphalan, mepacrine, mercaptopurine, methotrexate,metronidazole, mitomycin C, mitopodozide, mitotane, mitoxantrone,mithramycin, nalidixic acid, nifuratel, nifuroxazide, nifuralazine,nifurtimox, nimustine, ninorazole, nitrofurantoin, nitrogen mustards,oleomucin, oxolinic acid, pentamidine, pentostatin, phenazopyridine,phthalylsulfathiazole, pipobroman, prednimustine, prednisone, preussin,procarbazine, pyrimethamine, raltitrexed, rapamycin, rofecoxib,rosiglitazone, salazosulfapyridine, scriflavinium chloride, semustinestreptozocine, sulfacarbamide, sulfacetamide, sulfachlopyridazine,sulfadiazine, sulfadicramide, sulfadimethoxine, sulfaethidole,sulfafurazole, sulfaguanidine, sulfaguanole, sulfamethizole,sulfamethoxazole, co-trimoxazole, sulfamethoxydiazine,sulfamethoxypyridazine, sulfamoxole, sulfanilamide, sulfaperin,sulfaphenazole, sulfathiazole, sulfisomidine, staurosporin, tamoxifen,taxol, teniposide, tertiposide, testolactone, testosteronpropionate,thioguanine, thiotepa, timidazole, topotecan, triaziquone, treosulfan,trimethoprim, trofosfamide, UCN-01, vinblastine, vincristine, vindesine,vinblastine, vinorelbine, and zorubicin, or their respective derivativesor analogs thereof. Several of the above indicated drugs are nowadministered simultaneously for cancer therapy and, consequently, it isalso envisioned that more than one cytostatic and/or cytotoxic drug canbe comprised in compositions of the present invention.

As mentioned above, HIF inhibitors render cancer cells more vulnerableto chemotherapy and radiation therapy. Thus, to effectively treat ahyperproliferative disease or disorder, the compounds of the presentinvention can be co-administered with other active medicinal agentsand/or administered in conjunction with other anticancer, antitumor, orantiproliferative disease therapies. In one aspect, the inventionprovides a method for treating a hyperproliferative disease or disordercomprising administering a compound according to the invention to apatient prior to, during and/or after said patient was subjected to aradiation therapy, a chemotherapy, an immunotherapy, a laser/microwavethermotherapy or a gene therapy using antisense DNA and RNA (forexamples see Moeller et al., Cancer Cell 2004 5429-441).

In a further aspect the invention provides, as already outlined above,the use of a compound according to the invention or a compositionaccording to the invention for the preparation of a medicament for thetreatment or prevention of a disease or disorder selected from the groupconsisting of an inflammatory disease, a hyperproliferative disease ordisorder, a hypoxia related pathology such as e.g. diabetic retinopathy,ischemic reperfusion injury, ischemic myocardial and limb disease,ischemic stroke, sepsis and septic shock (see, e.g. Liu F Q, et al., ExpCell Res. 2008 Apr. 1; 314(6):1327-36); and a disease characterized bypathophysiological hyper-vascularisation, such as e.g. angiogenesis inosteosarcoma (see, e.g.: Yang, Qing-cheng et al., Dier Junyi DaxueXuebao (2008), 29(5), 504-508), macular degeneration, in particular,age-related macular degeneration and vasoproliferative retinopathy (seee.g. Kim J H, et al., J Cell Mol. Med. 2008 Jan. 19).

As was already mentioned above, HIF inhibitors, such as the compounds ofthe invention, are useful to treat inflammatory disease or disorder. Forexample, it was shown that oxygen-dependent HIF isoforms are stronglyupregulated in psoriatic skin (see e.g. Rosenberger C, et al., InvestDermatol. 2007 October; 127(10):2445-52). Furthermore it was shown thata HIF inhibitor, neovastat, inhibits the airway inflammation in asthma(see e.g., Lee S Y, et al., Vascul Pharmacol. 2007 November-December;47(5-6):313-8). Furthermore, recent evidence also shows that HIFparticipates under hypoxic conditions in joint inflammation anddestruction in rheumatoid arthritis (see e.g., Alin, J. K., et al.,Rheumatology (Oxford, United Kingdom) (2008), 47(6), 834-839). Thus, ina preferred embodiment of the use of the invention, the inflammatorydisease is selected form the group consisting of atherosclerosis,rheumatoid arthritis, asthma, inflammatory bowel disease, psoriasis, inparticular psoriasis vulgaris, psoriasis capitis, psoriasis guttata,psoriasis inversa; neurodermatitis; ichtyosises; alopecia greata;alopecia totalis; alopecia subtotalis; alopecia universalis; alopeciadiffusa; atopic dermatitis; lupus erythematodes of the skin;dermatomyositis of the skin; atopic eczema; morphea; scleroderma;alopecia greata Ophiasis type; androgenic alopecia; allergic dermatitis;irritative contact dermatitis; contact dermatitis; pemphigus vulgaris;pemphigus foliaceus; pemphigus vegetans; scarring mucous membranepemphigoid; bullous pemphigoid; mucous membrane pemphigoid; dermatitis;dermatitis herpetiformis Duhring; urticaria; necrobiosis lipoidica;erythema nodosum; prurigo simplex; prurigo nodularis; prurigo acuta;linear IgA dermatosis; polymorphic light dermatosis; erythema solaris;exanthema of the skin; drug exanthema; purpura chronica progressiva;dihydrotic eczema; eczema; fixed drug exanthema; photoallergic skinreaction; and periorale dermatitis. Therefore, a further preferredembodiment of the present invention encompasses a combination of one ormore compounds of the present invention and medication in current usefor treating such inflammatory diseases or conditions, which can bedetermined by a person skilled in the art of pharmacological sciences.Such therapeutics for combination can be selected e.g. from a group ofanti-inflammatory steroids, antioxidants, therapeutic antibodies orfusion proteins that sequester or bind to certain cytokines or cellularepitopes associated with inflammatory processes, or a dihydrofolatereductase inhibitor like methotrexate.

The compounds of the invention show anti-proliferative effects.Furthermore, HIF inhibitors, such as the compounds of the invention areeffective medicaments for the treatment of various cancer diseases (seereview article by e.g. Gregg L. Semenza, Nature Reviews, October 2003,vol. 3 and also review article by N. J. Mabjeesh et al., Histol.Histopathol (2007), 22:559-572). Thus, also preferred is the use of theinvention wherein the hyperproliferative disease is selected from thegroup consisting of a tumour or cancer disease, precancerosis,dysplasia, histiocytosis, a vascular proliferative disease and avirus-induced proliferative disease. Thus, in one preferred embodimentof the use of the invention the hyperproliferative disease is a tumor orcancer disease selected from the group consisting of diffuse largeB-cell lymphoma (DLBCL), T-cell lymphomas or leukemias, e.g., cutaneousT-cell lymphoma (CTCL), noncutaneous peripheral T-cell lymphoma,lymphoma associated with human T-cell lymphotrophic virus (HTLV), adultT- cell leukemia/lymphoma (ATLL), as well as acute lymphocytic leukemia,acute nonlymphocytic leukemia, acute myeloid leukemia, chroniclymphocytic leukemia, chronic myelogenous leukemia, Hodgkin's disease,non-Hodgkin's lymphoma, myeloma, multiple myeloma, mesothelioma,childhood solid tumors, glioma, bone cancer and soft-tissue sarcomas,common solid tumors of adults such as head and neck cancers (e.g., oral,laryngeal and esophageal), genitourinary cancers (e.g., prostate,bladder, renal (in particular malignant renal cell carcinoma (RCC)),uterine, ovarian, testicular, rectal, and colon), lung cancer (e.g.,small cell carcinoma and non-small cell lung carcinoma, includingsquamous cell carcinoma and adenocarcinoma), breast cancer, pancreaticcancer, melanoma and other skin cancers, basal cell carcinoma,metastatic skin carcinoma, squamous cell carcinoma of both ulceratingand papillary type, stomach cancer, brain cancer, liver cancer, adrenalcancer, kidney cancer, thyroid cancer, medullary carcinoma,osteosarcoma, soft-tissue sarcoma, Ewing's sarcoma, veticulum cellsarcoma, and Kaposi's sarcoma, fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, leiomyosarcoma, rhabdomyosarcoma, squamous cellcarcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma,sebaceous gland carcinoma, papillary carcinoma, papillaryadenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogeniccarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, small cell lungcarcinoma, epithelial carcinoma, astrocytoma, medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, meningioma, neuroblastoma, retinoblastoma,glaucoma, hemangioma, heavy chain disease and metastases.

The precancerosis treatable with the compounds of the present inventionare preferably selected from the group consisting of precancerosis, inparticular actinic keratosis, cutaneaous horn, actinic cheilitis, tarkeratosis, arsenic keratosis, x-ray keratosis, Bowen's disease, bowenoidpapulosis, lentigo maligna, lichen sclerosus, and lichen rubber mucosae;precancerosis of the digestive tract, in particular erythroplakia,leukoplakia, Barrett's esophagus, Plummer-Vinson syndrome, crural ulcer,gastropathia hypertrophica gigantea, borderline carcinoma, neoplasticintestinal polyp, rectal polyp, porcelain gallbladder; gynaecologicalprecancerosis, in particular carcinoma ductale in situ (CDIS), cervicalintraepithelial neoplasia (CIN), leukoplakia, endometrial hyperplasia(grade III), vulvar dystrophy, vulvar intraepithelial neoplasia (VIN),hydatidiform mole; urologic precancerosis, in particular bladderpapillomatosis, Queyrat's erythroplasia, testicular intraepithelialneoplasia (TIN), leukoplakia; carcinoma in situ (CIS); precancerosiscaused by chronic inflammation, in particular pyoderma, osteomyelitis,acne conglobata, lupus vulgaris, and fistula.

Dysplasia is frequently a forerunner of cancer, and is can be found ine.g. the epithelia; it is the most disorderly form of non-neoplasticcell growth, involving a loss in individual cell uniformity and in thearchitectural orientation of cells. Dysplastic cells often haveabnormally large, deeply stained nuclei, and exhibit pleomorphism.Dysplasia characteristically occurs where there exists chronicirritation or inflammation. Dysplastic disorders which can be treatedwith the compounds of the present invention include, but are not limitedto, anhidrotic ectodermal dysplasia, anterofacial dysplasia,asphyxiating thoracic dysplasia, atriodigital dysplasia,bronchopulmonary dysplasia, cerebral dysplasia, cervical dysplasia,chondroectodermal dysplasia, cleidocranial dysplasia, congenitalectodermal dysplasia, craniodiaphysial dysplasia, craniocarpotarsaldysplasia, craniometaphysial dysplasia, dentin dysplasia, diaphysialdysplasia, ectodermal dysplasia, enamel dysplasia, encephalo-ophthalmicdysplasia, dysplasia epiphysialis heminelia, dysplasia epiphysialismultiplex, dysplasia epiphysalis punctata, epithelial dysplasia,faciodigitogenital dysplasia, familial fibrous dysplasia of jaws,familial white folded dysplasia, fibromuscular dysplasia, fibrousdysplasia of bone, florid osseous dysplasia, hereditary renal-retinaldysplasia hidrotic ectodermal dysplasia, hypohidrotic ectodermaldysplasia, lymphopenic thymic dysplasia, mammary dysplasia,mandibulofacial dysplasia, metaphysical dysplasia, Mondini dysplasia,monostotic fibrous dysplasia, mucoepithelial dysplasia, multipleepiphysial dysplasia, oculoauriculovertebral dysplasia,oculodentodigital dysplasia, oculovertebral dysplasia, odontogenicdysplasia, ophthalmomandibulomelic dysplasia, periapical cementaldysplasia, polyostotic fibrous dysplasia, pseudoachondroplasticspondyloepiphysial dysplasia, retinal dysplasia, septo-optic dysplasia,spondyloepiphysial dysplasia, and ventriculoradial dysplasia.

Estrogen receptor refers to a group of receptors which are activated bythe hormone 17β-estradiol (estrogen). Two types of estrogen receptorexist: ER which is a member of the nuclear hormone family ofintracellular receptors and the estrogen G protein coupled receptorGPR30 (GPER), which is a G-protein coupled receptor. Estrogen and theestrogen receptors have been implicated in breast cancer, ovariancancer, colon cancer, prostate cancer and endometrial cancer and otherdiseases. As the compounds of the invention are capable of inhibitingestrogen receptor-mediated transcriptional activity, they can be used totreat said diseases.

Thus, in a further preferred embodiment, the hyperproliferativedisorders treatable according to the invention are those which benefitfrom a reduced estrogen receptor signalling, i.e. disorders associatedwith an increased estrogen receptor signaling, if compared to healthytissue. This particular suitability of the compounds of the presentinvention is based on the fact, that the compounds of the presentinvention potentially through inhibiting cellular replication butpossibly also through an additional activity of the compounds of thepresent invention exert an inhibition of estrogen receptor signaling.Thus, preferred diseases, conditions and/or disorders which can betreated are selected from the group consisting of mammary tumors,endometrial tumors and tumors of the uterus. Whether a disease isassociated an increased estrogen receptor activity can be measured by avariety of art known methods including determination of ER expressionlevel in the diseased tissue by, e.g. immunological methods, whichdetermine the amount of expressed protein, by methods determining theamount of transcribed ER encoding nucleic acids, e.g. RT-PCR,Northern-blots, nuclear run-ons etc., and determining the activity of anucleic acid construct comprising an ER-receptor recognition element,which drives expression of a detectable reporter, e.g. CAT, luciferase,GFP etc as described in more detail in the Experimental Section below.Preferably, the disorders which benefit from a reduced estrogen receptorsignaling are those, which show in the diseased tisse an increase inestrogen receptor signaling by at least 10%, preferably by at least 20%,30%, 40%, 50%, 60%, 70%, if compared to healthy tissue. Preferably thisincrease is measured on the basis of a nucleic acid comprising anER-receptor recognition element and the increase of the expression of areporter driven by this element.

In therapeutic use as an antagonist of estrogen receptor signaling,acting through inhibition of cellular replication, the compoundsutilized in the use of the invention are administered at the initialdosage of about 0.02 mg/kg to about 20 mg/kg daily. A daily dose rangeof about 0.05 mg/kg to about 2 mg/kg is preferred, with a daily doserange of about 0.05 mg/kg to about 1 mg/kg being most preferred. Thedosages, however, may be varied depending upon the requirements of thepatient, the severity of the condition being treated, and the compoundbeing employed. Determination of the proper dosage for a particularsituation is within the skill of the practitioner. Generally, treatmentis initiated with smaller dosages, which are less than the optimum doseof the compound. Thereafter, the dosage is increased by small incrementsuntil the optimum effect under circumstances is reached. Forconvenience, the total daily dosage may be divided and administered inportions during the day, if desired.

Salts/Esters

The compounds within the compositions or compounds usable according tothe present invention can be present as salts or esters, in particularpharmaceutically acceptable salts or esters. Pharmaceutically acceptablesalts of the compounds of the invention include suitable acid additionor base salts thereof A review of suitable pharmaceutical salts may befound in Berge et al, J Pharm Sci, 66, 1-19 (1977). Salts are formed,for example with strong inorganic acids such as mineral acids, e.g.sulphuric acid, phosphoric acid or hydrohalic acids; with strong organiccarboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atomswhich are unsubstituted or substituted (e.g., by halogen), such asacetic acid; with saturated or unsaturated dicarboxylic acids, forexample oxalic, malonic, succinic, maleic, fumaric, phthalic ortetraphthalic; with hydroxycarboxylic acids, for example ascorbic,glycolic, lactic, malic, tartaric or citric acid; with aminoacids, forexample aspartic or glutamic acid; with benzoic acid; or with organicsulfonic acids, such as (C1-C4)-alkyl- or aryl-sulfonic acids which areunsubstituted or substituted (for example, by a halogen) such asmethane- or p-toluene sulfonic acid.

Esters are formed either using organic acids or alcohols/hydroxides,depending on the functional group being esterified. Organic acidsinclude carboxylic acids, such as alkanecarboxylic acids of 1 to 12carbon atoms which are unsubstituted or substituted (e.g., by halogen),such as acetic acid; with saturated or unsaturated dicarboxylic acid,for example oxalic, malonic, succinic, maleic, fumaric, phthalic ortetraphthalic; with hydroxycarboxylic acids, for example ascorbic,glycolic, lactic, malic, tartaric or citric acid; with aminoacids, forexample aspartic or glutamic acid; with benzoic acid; or with organicsulfonic acids, such as (C1-C4)-alkyl- or aryl-sulfonic acids which areunsubstituted or substituted (for example, by a halogen) such asmethane- or p-toluene sulfonic acid. Suitable hydroxides includeinorganic hydroxides, such as sodium hydroxide, potassium hydroxide,calcium hydroxide, aluminium hydroxide. Alcohols include alkanealcoholsof 1-12 carbon atoms which may be unsubstituted or substituted, e.g. bya halogen).

Isotopes

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. An isotopic variation of an agent of thepresent invention or a pharmaceutically acceptable salt thereof isdefined as one in which at least one atom is replaced by an atom havingthe same atomic number but an atomic mass different from the atomic massusually found in nature. Examples of isotopes that can be incorporatedinto the agent and pharmaceutically acceptable salts thereof includeisotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulphur,fluorine and chlorine such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³¹P, ³²P,³⁵S, ¹⁸F and ³⁶Cl, respectively. Certain isotopic variations of theagent and pharmaceutically acceptable salts thereof, for example, thosein which a radioactive isotope such as ³H or ¹⁴C is incorporated, areuseful in drug and/or substrate tissue distribution studies. Tritiated,i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes are particularly preferredfor their ease of preparation and detectability. Further, substitutionwith isotopes such as deuterium, i.e., ³H, may afford certaintherapeutic advantages resulting from greater metabolic stability, forexample, increased in vivo half-life or reduced dosage requirements andhence may be preferred in some circumstances. Isotopic variations of theagent of the present invention and pharmaceutically acceptable saltsthereof of this invention can generally be prepared by conventionalprocedures using appropriate isotopic variations of suitable reagents.

All isotopic variations of the compounds and compositions of the presentinvention, whether radioactive or not, are intended to be encompassedwithin the scope of the present invention.

Solvates

The present invention also includes solvate forms of the compoundswithin the compositions or compounds according to any of generalformulas (I) through (III) usable according to the present invention.The terms used in the claims encompass these forms.

Polymorphs

The invention furthermore relates to compounds within the compositionsof the present invention or compounds according to formula (I) usableaccording to the present invention in their various crystalline forms,polymorphic forms and (an)hydrous forms. It is well established withinthe pharmaceutical industry that chemical compounds may be isolated inany of such forms by slightly varying the method of purification and orisolation form the solvents used in the synthetic preparation of suchcompounds.

Administration

A compound according to the invention can be administered by variouswell known routes, including oral, rectal, intragastrical, intracranialand parenteral administration, e.g. intravenous, intramuscular,intranasal, intradermal, subcutaneous, and similar administrationroutes. Parenteral administration and particular intravenousadministration, preferably by depot injection, is preferred. Dependingon the route of administration different pharmaceutical formulations arerequired and some of those may require that protective coatings areapplied to the drug formulation to prevent degradation of a compound ofthe invention in, for example, the digestive tract.

Thus, preferably, a compound of the invention is formulated as a syrup,an infusion or injection solution, a tablet, a capsule, a capslet,lozenge, a liposome, a suppository, a plaster, a band-aid, a retardcapsule, a powder, or a slow release formulation. Preferably the diluentis water, a buffer, a buffered salt solution or a salt solution and thecarrier preferably is selected from the group consisting of cocoa butterand vitebesole.

Particular preferred pharmaceutical forms for the administration of acompound of the invention are forms suitable for injectionable use andinclude sterile aqueous solutions or dispersions and sterile powders forthe extemporaneous preparation of sterile injectable solutions ordispersion. In all cases the final solution or dispersion form must besterile and fluid. Typically, such a solution or dispersion will includea solvent or dispersion medium, containing, for example, water-bufferedaqueous solutions, e.g. biocompatible buffers, ethanol, polyol, such asglycerol, propylene glycol, polyethylene glycol, suitable mixturesthereof, surfactants or vegetable oils. A compound of the invention canalso be formulated into liposomes, in particular for parenteraladministration. Liposomes provide the advantage of increased half lifein the circulation, if compared to the free drug and a prolonged moreeven release of the enclosed drug.

Sterilization of infusion or injection solutions can be accomplished byany number of art recognized techniques including but not limited toaddition of preservatives like anti-bacterial or anti-fungal agents,e.g. parabene, chlorobutanol, phenol, sorbic acid or thimersal. Further,isotonic agents, such as sugars or salts, in particular sodium chloridemay be incorporated in infusion or injection solutions.

Production of sterile injectable solutions containing one or several ofthe compounds of the invention is accomplished by incorporating therespective compound in the required amount in the appropriate solventwith various ingredients enumerated above as required followed bysterilization. To obtain a sterile powder the above solutions arevacuum-dried or freeze-dried as necessary. Preferred diluents of thepresent invention are water, physiological acceptable buffers,physiological acceptable buffer salt solutions or salt solutions.Preferred carriers are cocoa butter and vitebesole. Besides thepreferred excipients mentioned already above, also the followingexcipients can be chosen, without limitation, to be used with thevarious pharmaceutical forms of a compound of the invention:

-   a) binders such as lactose, mannitol, crystalline sorbitol, dibasic    phosphates, calcium phosphates, sugars, microcrystalline cellulose,    carboxymethyl cellulose, hydroxyethyl cellulose, polyvinyl    pyrrolidone and the like;-   b) lubricants such as magnesium stearate, talc, calcium stearate,    zinc stearate, stearic acid, hydrogenated vegetable oil, leucine,    glycerids and sodium stearyl fumarates,-   c) disintegrants such as starches, croscaramellose, sodium methyl    cellulose, agar, bentonite, alginic acid, carboxymethyl cellulose,    polyvinyl pyrrolidone and the like.

Other suitable excipients can be found in the Handbook of PharmaceuticalExcipients, published by the American Pharmaceutical Association, whichis herein incorporated by reference.

It is to be understood that depending on the severity of the disorderand the particular type which is treatable with one of the compounds ofthe invention, as well as on the respective patient to be treated, e.g.the general health status of the patient, etc., different doses of therespective compound are required to elicit a therapeutic or prophylacticeffect. The determination of the appropriate dose lies within thediscretion of the attending physician. It is contemplated that theaverage daily dosage of a compound of the invention in the therapeuticor prophylactic use of the invention should be in the range of about 0.1mg to about 3 g. However, in a preferred use of the present invention acompound of the invention is administered to a subject in need thereofin an amount ranging from 1.0 to 1000 mg, preferably ranging from 10 to500 mg preferably ranging from 50 to 200 mg. The duration of therapy andthe dosing frequency with a compound of the invention will vary,depending on the severity of the disease being treated and the conditionand idiosyncratic response of each individual patient.

As is known in the art, the pharmaceutically effective amount of a givencomposition will also depend on the administration route. In general therequired amount will be higher, if the administration is through thegastrointestinal tract; e.g. by suppository, rectal, or by anintragastric probe, and lower if the route of administration isparenteral, e.g. intravenous. Typically, a compound of the inventionwill be administered in ranges of 50 mg to 3 g, preferably 50 mg to 500mg, if rectal or intragastric administration is used and in ranges of 10to 500 mg, if parenteral administration is used.

If a person is know to be at risk of developing a disorder treatablewith a compound of the invention, a prophylactic administration of thepharmaceutical composition according to the invention may be possible.In these cases, the respective compound of the invention is preferablyadministered in above outlined preferred and particular preferred doseson a daily basis. This administration can be continued until the risk ofdeveloping the respective disorder has lessened. In most instances,however, a compound of the invention will be administered once adisease/disorder has been diagnosed. In these cases it is preferred thata first dose of a compound of the invention is administered one, two,three or four times daily. Preferably the administration is discontinuedfor one day, one week or one month and then repeated until the symptomsof the respective disease are no longer worsening or until they areimproving.

Within the meaning of this invention, a combination of substituents orvariables is permissible only if such a combination results in a stableor chemically feasible compound. A stable compound or chemicallyfeasible compound is one that is not substantially altered when kept ata temperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week. This invention alsoenvisions the quaternization of any basic nitrogen-containing groups ofthe compounds disclosed herein. Water or oil-soluble or dispersibleproducts may be obtained by such quaternization.

Various modifications and variations of the invention will be apparentto those skilled in the art without departing from the scope of theinvention. Although the invention has been described in connection withspecific preferred embodiments, it should be understood that theinvention as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the invention which are obvious to those skilled in therelevant fields are intended to be covered by the present invention.

The following examples and figures are merely illustrative of thepresent invention and should not be construed to limit the scope of theinvention as indicated by the appended claims in any way.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1: Depicts the fluorescence-assisted cell sorting (FACS) dataobtained in example 8 described below.

EXPERIMENTAL SECTION

All starting materials used in the examples are either commerciallyavailable or can be synthesized by the average skilled person trained inorganic chemistry without undue burden following routine laboratorypractice as outlined, for example in example 1.

Besides the guidance provided in example 1, alternative synthetictransformations that may be employed in the synthesis of compounds offormulas I-III and in the synthesis of intermediates involved in thesynthesis of compounds of formulas I-III are known by or accessible toone skilled in the art. Collections of synthetic transformations may befound in compilations, such as: J. March. Advanced Organic Chemistry,4th ed.; John Wiley: New York (1992) R. C. Larock. Comprehensive OrganicTransformations, 2nd ed.; Wiley-VCH: New York (1999); F. A. Carey; R. J.Sundberg. Advanced Organic Chemistry, 2nd ed.; Plenum Press: New York(1984) T. W. Greene; P. G. M. Wuts. Protective Groups in OrganicSynthesis, 3rd ed.; John Wiley: New York (1999). L. S. Hegedus.Transition Metals in the Synthesis of Complex Organic Molecules, 2nded.; University Science Books: Mill Valley, Calif. (1994) L. A.Paquette, Ed. The Encyclopedia of Reagents for Organic Synthesis; JohnWiley: New York (1994). A. R. Katritzky; 0. Meth-Cohn; C. W. Rees, Eds.Comprehensive Organic Functional Group Transformations; Pergamon Press:Oxford, UK (1995). G. Wilkinson; F. G A. Stone; E. W. Abel, Eds.Comprehensive Organometallic Chemistry; Pergamon Press: Oxford, UK(1982). B. M. Trost; I. Fleming. Comprehensive Organic Synthesis;Pergamon Press: Oxford, UK (1991) A. R. Katritzky; C. W. Rees Eds.Comprehensive Heterocylic Chemistry; Pergamon Press: Oxford, UK (1984)A. R. Katritzky; C. W. Rees; E. F. V. Scriven, Eds. ComprehensiveHeterocylic Chemistry; Pergamon Press: Oxford, UK (1996). C. Hansch; P.G. Sammes; J. B. Taylor, Eds. Comprehensive Medicinal Chemistry:Pergamon Press: Oxford, UK (1990).

In addition, recurring reviews of synthetic methodology and relatedtopics include Organic Reactions; John Wiley: New York; OrganicSyntheses; John Wiley: New York; Reagents for Organic Synthesis: JohnWiley: New York; The Total Synthesis of Natural Products; John Wiley:New York; The Organic Chemistry of Drug Synthesis; John Wiley: New York;Annual Reports in Organic Synthesis; Academic Press: San Diego Calif.;and Methoden der Organischen Chemie (Houben-Weyl); Thieme: Stuttgart,Germany. Furthermore, databases of synthetic transformations includeChemical Abstracts, which may be searched using either CAS OnLine orSciFinder, Handbuch der Organischen Chemie (Beilstein), which may besearched using SpotFire, and REACCS.

The compounds herein were named according to IUPAC standard using thesoftware AutoNom Standard For ISIS/Draw Add-In.

Example 1 Synthesis of the Compounds of the Invention

The compounds according to general formulas (I) through (III) accordingto the present invention may be prepared for example according to thefollowing scheme:

wherein R¹ through R⁶ are as defined in the claims, or have theparticularly preferred meanings as defined herein and Rx is H and/orpinacolato.

The general reaction schemes indicated above are as follows:

General Procedure A. A flask charged with 1 (1.0 equiv),Pd(dppf)Cl2.CH₂Cl₂ (0.1 equiv), and the corresponding heteroaromatic oraromoatic boronic acid or boronic ester (1.2 equiv) in 1:1 (v/v)Toluene/MeOH was purged with N₂ under vigorous stirring, and 2.0 Maqueous K₂CO₃ solution (2.5 equiv) were added slowly. The mixture washeated at 90° C. for 4 h. After cooling, the mixture was purified byflash column chromatography (silica gel, EtOAc/heptane, 1:5 to 1:3) togive the derivatives 2 (43-68% yield).

General Procedure B. A flask charged with aniline derivatives 1 or 2(1.0 equiv), the corresponding sulfonyl chloride (1.1 equiv) andpyridine (10 equiv) in CH₂Cl₂ was stirred at room temperature for 12 h.The mixture was extracted with concentrated aqueous NH₄Cl, washed withbrine and dried over MgSO₄. The organic solvents were evaporated and thecrude product was purified by preparative HPLC or by flash columnchromatography (silica gel, EtOAc/heptane) to give the derivatives 3 or4 (70% to quantitative yield).

General Procedure C. A 2-5 ml microwave reaction vessel charged with 4(1.0 equiv), Pd(dppf)Cl₂.CH₂Cl₂ (0.1 equiv), and the correspondingheteroaromatic or aromatic boronic acid or boronic ester (1 equiv) indry MeOH was purged with N₂ under vigorous stirring and 2.0 M aqueousK₂CO₃ solution (2.5 equiv) were added slowly. The vessel was sealed andthe mixture was heated in a microwave oven for 30 min at 90° C. (CEMDiscover Microwave system, set to P_(max)=150W). After cooling, themixture was diluted with CH₂Cl₂, filtered through celite and eluted withCH₂Cl₂. The organic solvents were evaporated and the crude product waspurified by preparative HPLC or by flash column chromatography (silicagel, EtOAc/heptane,) to give the derivatives 3 (42-90% yield).

General Procedure D. Approach 2 can optionally include an extra step asshown. Bispinacolato diboron (2.5 equiv), Pd(dppf)Cl₂.CH₂Cl₂ (0.1equiv), and KOAc (3.0 equiv) were added to a flask containing compounds4 (1.0 equiv) in degassed 1,4-dioxane. The reaction mixture was heatedto 95° C. and stirred for 4 hours. Upon completion, the reaction mixturewas diluted with EtOAc (50 ml), filtered through a short column ofsilica gel, and further eluted with EtOAc. The combined organic solventwas washed with H₂O and brine, dried over anhydrous MgSO₄ andconcentrated in vacuum. The resulting residue was purified by flashcolumn chromatography (silica gel, EtOAc/heptane, 1:3) to yield theboronate esters 5 (50-88% yield) as a cream or white solid. In asubsequent step, compounds 5 were transformed with a heteroaromatic oraromatic bromide to 3 following procedures A or C. Substituents R¹, R⁴and/or R⁶, which may undergo unwanted reactions when carrying out asynthesis according to approach 1 or 2 may be protected by aconventional protecting group, which is not cleaved during the reactionsaccording to approach 1 or 2 but is cleavable under known conditions.The skilled person is aware of a large variety of protection groups,which can be employed in organic synthesis. Protecting groups arereviewed in, for example, Wuts, P. G. M. and Greene, T. W., ProtectiveGroups in Organic Chemistry, 3^(rd) Ed., 1999; Wily & Sons Inc. and inKocienski, P. J., Protecting groups; 2″ Ed., 2000, Thieme MedicalPublishing. Protecting groups are organized in these reference booksaccording to the functionalities that are protected as well as accordingto the conditions which remove the respective protecting groupsselectively. Particularly preferred protective groups, which can beused, are:

-   (i) protective group removed at acidic conditions, preferably at a    pH between 4 and 6, which is selected from the group consisting of    Boc or Trityl protecting groups;-   (ii) a protecting group removed by a nucleophile, which is selected    from the group consisting of Fmoc or Dde protecting groups;-   (iii) a protecting group removed by hydrogenolysis consisting of the    allyl type, the tert-butyl type, the benzyl type or Dmab    (4-{N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino}benzyl    ester);-   (iv) a protecting group removed by radiation, which is selected from    the group consisting of nitroveratryloxy carbonyl, nitrobenzyloxy    carbonyl, dimethyl dimethoxybenzyloxy carbonyl,    5-bromo-7-nitroindolinyl, o-hydroxy-α-methyl cinnamoyl, and    2-oxymethylene anthraquinone.

Synthesis of Building Blocks

The toluidine building blocks (1) are commercially available, but canalso be synthesized according to standard functionalization ortransformation protocols, described in the literature cited above, andnormally known to those ordinary skilled in the art of organicsynthesis.

Numerous aromatic or heteroaromatic boronic acids or esters useful inProcedures A and C for palladium catalyzed coupling reactions arecommercially available, or they can be synthesized according to standardfunctionalization or transformation protocols, described in theliterature cited above, and normally known to those ordinary skilled inthe art of organic synthesis. In particular, such boronic acid/esterintermediates can be generated e.g. from their corresponding aromatic orheteroaromatic halide precursors, as exemplified in Procedure D.

Numerous 5- and 6-membered heteroaromatic sulfonic acids and theircorresponding activated derivatives, as used in Procedure B and thereexemplified for sulfonylchlorides, are commercially available or theycan be synthesized according to standard functionalization ortransformation protocols, described in the literature cited above andnormally known to those ordinary skilled in the art of organicsynthesis. (Particular methods are e.g. described in: Caldwell, W T etal., J. Med. Chem. (1962), Vol. 6, p. 58 ff; Caldwell, W T et al., JACS(1959), Vol. 81, p. 5166 ff; Roblin, R O et al., JACS (1950), Vol. 72,p. 4890 ff; Hitoshi K et al., US Patent (1998), U.S. Pat. No. 5,811,571;WO 2006/090244, p. 38 ff; Janosik T et al., THL (2006), Vol. 62, p. 1699ff; Allred GD, SERMACS (2007), Lanny Liebeskind Cope Scholar AwardSymposium II, 222. Sulfonyl Fluorides; and references cited therein). Ifdesired, it is also possible to generate sulfonamides 3 and/or 4directly from the corresponding thiols (Wright S W et al., JOC (2006),Vol. 71, p. 1080 ff).

Example 2 HPLC/MS Analysis of the Compounds of the Invention

The compounds were analyzed as follows:

Measured via HPLC/MS, using a Waters X-bridge C₁₈-column, 5 μm particlesize, 4.6×150 mm (diameter×length) at a flow rate of 1.75 ml/min with alinear gradient (water to acetonitrile, 0.2% formic acid as modifier)from initially 99:1 to 1:99 over 9.10 min, then hold for 1.80 min. Masssignals were determined using a Waters 3100 Mass Detector.

Example 3 General Cell Culture Maintenance and Cell Proliferation Assays

MCF-7 human breast adenocarcinoma cells and HL-60 acute promyelocyticleukaemia cells were obtained from ATCC (LGC Promochem). HG-1 multiplemyeloma cells were obtained from Dr. D. Hose (DKFZ Heidelberg). TheCellSensor® HRE-bla HCT116 cell line (colorectal carcinoma) was obtainedfrom Invitrogen.

Cells were grown under humidified 95% air, 5% CO₂ at 37° C. inDulbecco's modified Eagle's medium (DMEM: MCF-7) or RPMI 1640 medium(RPMI: HL-60, HG-1) supplemented with 10% fetal bovine serum (FBS), 100U/ml penicillin and 100 μg/ml streptomycin, 2 mM L-glutamine, and 2ng/ml IL-6 (for the HG-1 cell line only). The HCT116 cell line was grownunder the same atmospheric conditions in McCoy's 5A medium supplementedwith 10% FBS and penicillin/streptomycin as above, in addition toblasticidin as a selection marker at a final concentration of 5 μg/ml.

Cell proliferation experiments were carried out in 96-well tissueculture plates with seeding of 2000 cells/well (MCF-7) or 1000cells/well (HL-60, HG-1) in 100 μl of the relevant medium. Cells weresubsequently incubated under the conditions mentioned for 24 h prior toaddition of compounds.

For the determination of compound EC₅₀ values, 10 μl compounds at 11×concentrations in 5.5% DMSO were added to the wells at variousconcentrations yielding a final constant percentage of 0.5% DMSO at thedesired 1× compound concentration. As a positive control, cells weretreated with 5.5% DMSO. The cells were then incubated for a further 72 hprior to the measurement.

To determine the degree of inhibition of cell proliferation, cells weretreated with ATPlite solution according to the manufacturer'sinstructions (PerkinElmer, ATPlite 1-step Luminescence ATP DetectionAssay System) and luciferase readout measured on an Envision HTSmultilabel plate reader (PerkinElmer) in luminescence mode according toestablished protocols. Raw data were imported into an ActivityBasedatabase (IDBS, ID Business Solutions) and EC₅₀ values calculated usingthe IDBS program ActivityBase XE. Table 2A and 2B show the IC₅₀ valuesobtained in the above-outlined cell proliferation assay for exemplarycompounds of the invention. The cell proliferation assay data show thatthe compounds of the invention can inhibit cell proliferation in cancercell lines.

TABLE 2A MCF7 cells Mass Example Retention time (HPLC) Spectromety no.IUPAC Name EC₅₀ [min] MH+ 1 4-Methyl-N-[2-methyl-5-(1H-tetrazol- “+”5.87 330 5-yl)-phenyl]-benzenesulfonamide 24-Methyl-N-(2-methyl-5-tetrazol-1-yl- “+” 6.27 330phenyl)-benzenesulfonamide 3 4-Methyl-N-[2-methyl-5-(5-methyl-furan-“+++” 8.27 343 2-yl)-phenyl]-benzenesulfonamide 4N-(5-Furan-2-yl-2-methyl-phenyl)-4- “+++” 7.92 328methyl-benzenesulfonamide 5 N-(5-Furan-3-yl-2-methyl-phenyl)-4- “+++”7.73 328 methyl-benzenesulfonamide 64-Methyl-N-[2-methyl-5-(2-methyl-2H- “+” 6.88 344tetrazol-5-yl)-phenyl]-benzenesulfonamide 7N-(4,3′-Dimethyl-biphenyl-3-yl)-4-methyl- “++” 8.7 352benzenesulfonamide 8 4-Methyl-N-(2-methyl-5-thiophen-3-yl- “+++” 8.12344 phenyl)-benzenesulfonamide 9 N-[5-(6-Methoxy-pyridin-2-yl)-2-methyl-“++” 8.2 370 phenyl]-4-methyl-benzenesulfonamide 10N-(4,4′-Dimethyl-biphenyl-3-yl)-4-methyl- “+” 8.72 352benzenesulfonamide 11 4-Methyl-N-[2-methyl-5-(5-methyl-thiophen- “+++”8.65 358 2-yl)-phenyl]-benzenesulfonamide 12N-[5-(2,4-Dimethyl-thiazol-5-yl)-2- “+++” 7.07 373methyl-phenyl]-4-methyl- benzenesulfonamide 13N-[5-(2,5-Dimethyl-thiophen-3-yl)-2- “+” 8.63 388methyl-phenyl]-4-methoxy- benzenesulfonamide 144-Methoxy-N-(2-methyl-5-thiophen-3- “+++” 7.72 360yl-phenyl)-benzenesulfonamide 15 4-Methyl-N-(2-methyl-5-pyrrol-1-yl-“+++” 7.92 327 phenyl)-benzenesulfonamide 16N-[5-(3,5-Dimethyl-isoxazol-4-yl)-2- “+++” 7.03 373methyl-phenyl]-4-methoxy- benzenesulfonamide 174-Methoxy-N-[2-methyl-5-(2-methyl- “+++” 8.23 374 thiophen-3-yl)-phenyl]-benzenesulfonamide 18N-[5-(6-Methoxy-pyridin-3-yl)-2-methyl- “+” 7.73 368.94phenyl]-4-methyl-benzenesulfonamide 194-Methyl-N-(2-methyl-5-pyridin-2-yl- “++” 5.94 338, 340phenyl)-benzenesulfonamide 20 4-Methoxy-N-(2-methyl-5-oxazol-5-yl- “+++”6.36 345 phenyl)-benzenesulfonamide 214-Methoxy-N-(2-methyl-5-pyrrol-1-yl- “+++” n.d. n.d.phenyl)-benzenesulfonamide 22 4-Methyl-N-(2-methyl-5-[1,2,4]triazol- “+”5.26 329 4-yl-phenyl)-benzenesulfonamide 234-Methyl-N-[2-methyl-5-(4-methyl-thiophen- “+++” 8.46 3583-yl)-phenyl]-benzenesulfonamide 244-Methyl-N-[2-methyl-5-(3-methyl-thiophen- “+++” 8.53 3582-yl)-phenyl]-benzenesulfonamide 254-Methyl-N-[2-methyl-5-(2-methyl-furan- “+++” 8.11 3423-yl)-phenyl]-benzenesulfonamide 26N-[5-(3-Methoxy-pyridin-2-yl)-2-methyl- “+” 5.8 368.98phenyl]-4-methyl-benzenesulfonamide 274-Methyl-N-[2-methyl-5-(3-trifluoromethyl- “+” 7.55 407pyridin-2-yl)-phenyl]-benzenesulfonamide 284-Methyl-N-[2-methyl-5-(3-methyl-pyridin- “+” 4.95 3532-yl)-phenyl]-benzenesulfonamide 29N-[5-(2,5-Dimethyl-pyrrol-1-yl)-2-methyl- “+” 8.36 355phenyl]-4-methyl-benzenesulfonamide 304-Methoxy-N-[2-methyl-5-(3-methyl- “+++” 7.14 358pyrazol-1-yl)-phenyl]-benzenesulfonamide 314-Methyl-N-[2-methyl-5-(4-methyl-pyridin- “+” 5.28 352.982-yl)-phenyl]-benzenesulfonamide 324-Methyl-N-[2-methyl-5-(5-methyl-pyridin- “+” 5.88 3532-yl)-phenyl]-benzenesulfonamide 334-Methyl-N-[2-methyl-5-(6-methyl-pyridin- “++” 5.4 3532-yl)-phenyl]-benzenesulfonamide 34N-[5-(5-Cyano-pyridin-3-yl)-2-methyl- “+++” 7 364phenyl]-4-methyl-benzenesulfonamide 35N-[5-(5-Methoxy-pyridin-3-yl)-2-methyl- “+” 5.88 369phenyl]-4-methyl-benzenesulfonamide 36N-[5-(6-Methoxy-4-methyl-pyridin-3- “+” 7.83 382.99yl)-2-methyl-phenyl]-4-methyl- benzenesulfonamide 37N-[5-(6-Ethoxy-pyridin-3-yl)-2-methyl- “+” 8.18 383phenyl]-4-methyl-benzenesulfonamide 384-Methyl-N-[2-methyl-5-(4-trifluoromethyl- “+” 8.29 407pyridin-2-yl)-phenyl]-benzenesulfonamide 394-Methyl-N-[2-methyl-5-(6-trifluoromethyl- “++” 8.36 407pyridin-2-yl)-phenyl]-benzenesulfonamide 40N-[5-(2,4-Dimethyl-pyrrol-1-yl)-2-methyl- “+” n.d. n.d.phenyl]-4-methoxy-benzenesulfonamide 41N-[5-(2-Cyano-pyrrol-1-yl)-2-methyl- “++” 7.95* 368phenyl]-4-methoxy-benzenesulfonamide 42N-[5-(6-Acetyl-pyridin-2-yl)-2-methyl- “++” 7.7 381phenyl]-4-methyl-benzenesulfonamide 43N-[5-(5-Cyano-pyridin-2-yl)-2-methyl- “+++” 7.38 364phenyl]-4-methyl-benzenesulfonamide 44N-[5-(5-Methoxy-pyridin-2-yl)-2-methyl- “+++” 6.91 369phenyl]-4-methyl-benzenesulfonamide 454-Methyl-N-[2-methyl-5-(5-trifluoromethyl- “+” 8.39 407pyridin-2-yl)-phenyl]-benzenesulfonamide 46N-[5-(5-Fluoro-6-methyl-pyridin-2-yl)- “++” 8.12 3712-methyl-phenyl]-4-methyl- benzenesulfonamide 47N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “+++” 7.93 369phenyl]-4-methyl-benzenesulfonamide 48N-[5-(3,5-Dimethyl-isoxazol-4-yl)-2- “+++” 7.34 357methyl-phenyl]-4-methyl- benzenesulfonamide 49N-[5-(5-Cyano-pyridin-2-yl)-2-methyl- “+++” 7.06 380phenyl]-4-methoxy-benzenesulfonamide 504-Methoxy-N-[2-methyl-5-(2-methyl- “+++” 7.81 357pyrrol-1-yl)-phenyl]-benzenesulfonamide 51N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “+++” 7.64 385phenyl]-4-methoxy-benzenesulfonamide 52N-[5-(6-Cyano-pyridin-2-yl)-2-methyl- “++” 7.5 364phenyl]-4-methyl-benzenesulfonamide 53N-[5-(5-Fluoro-pyridin-2-yl)-2-methyl- “+++” 7.63 356.75phenyl]-4-methyl-benzenesulfonamide 54N-[5-(5-Acetyl-pyridin-2-yl)-2-methyl- “+” 7.07 381phenyl]-4-methyl-benzenesulfonamide 554-Methyl-N-[2-methyl-5-(5-nitro-pyridin- “+++” 7.71 3842-yl)-phenyl]-benzenesulfonamide 56N-[5-(4-Cyano-thiophen-2-yl)-2-methyl- “+++” 7.45 385phenyl]-4-methoxy-benzenesulfonamide 57N-(3′-Cyano-4-methyl-biphenyl-3-yl)- “++” 7.49 3794-methoxy-benzenesulfonamide 58 N-(2′-Cyano-4-methyl-biphenyl-3-yl)-“++” 7.38 379 4-methoxy-benzenesulfonamide 59N-[5-(3-Cyano-thiophen-2-yl)-2-methyl- “++” 7.67 369phenyl]-4-methyl-benzenesulfonamide 60N-(4′-Cyano-4-methyl-biphenyl-3-yl)- “+++” 7.83 3634-methyl-benzenesulfonamide 61 N-[5-(6-Cyano-pyridin-3-yl)-2-methyl-“+++” 7.02 380 phenyl]-4-methoxy-benzenesulfonamide 62N-[5-(2-Cyano-thiophen-3-yl)-2-methyl- “+++” 7.41 384phenyl]-4-methoxy-benzenesulfonamide 63N-(4′-Cyano-4-methyl-biphenyl-3-yl)- “+++” 7.53 3794-methoxy-benzenesulfonamide 64 N-[5-(5-Cyano-pyrimidin-2-yl)-2-methyl-“++” 7.27 365 phenyl]-4-methyl-benzenesulfonamide 65N-(3′,4′-Dicyano-4-methyl-biphenyl- “+++” 8.06 4043-yl)-4-methoxy-benzenesulfonamide 66N-[5-(6-Dimethylamino-pyridin-2-yl)- “++” 6.8 3832-methyl-phenyl]-4-methyl- benzenesulfonamide 67N-[5-(4,6-Dimethyl-pyridin-2-yl)-2- “+” 5.14 367methyl-phenyl]-4-methyl- benzenesulfonamide 68N-[5-(5-Cyano-thiophen-3-yl)-2-methyl- “+++” 7.9 369phenyl]-4-methyl-benzenesulfonamide 69N-[5-(4-Cyano-thiophen-3-yl)-2-methyl- “+++” 7.55 369phenyl]-4-methyl-benzenesulfonamide 70 N-[5-(3-Cyo-furan-2-yl)-2-methyl-“+++” 7.55 353 phenyl]-4-methyl-benzenesulfonamide 71N-(4′-Cyano-3′-fluoro-4-methyl-biphenyl- “+++” 8 3813-yl)-4-methyl-benzenesulfonamide 72 N-(4′-Cyano-4,3′-dimethyl-biphenyl-“+++” 8.13 377 3-yl)-4-methyl-benzenesulfonamide 73N-[5-(5-Cyano-thiophen-3-yl)-2-methyl- “+++” 7.53 385phenyl]-4-methoxy-benzenesulfonamide 74N-[5-(5-Cyano-pyridin-3-yl)-2-methyl- “+++” 5.98 338.92phenyl]-4-methoxy-benzenesulfonamide 75N-[5-(2-Cyano-pyridin-4-yl)-2-methyl- “++” 6.95 380phenyl]-4-methoxy-benzenesulfonamide 764-Methyl-N-[2-methyl-5-(5-nitro-furan- “+++” 7.62 3732-yl)-phenyl]-benzenesulfonamide 774-Chloro-N-[5-(5-cyano-thiophen-2-yl)- “+++” 8.26 3892-methyl-phenyl]-benzenesulfonamide 78N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “++” 8.4 423phenyl]-4-trifluoromethyl- benzenesulfonamide 79N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “+” 8.48 439phenyl]-4-trifluoromethoxy- benzenesulfonamide 80N-[5-(4-Cyano-thiophen-2-yl)-2-methyl- “+++” 8.21 385phenyl]-4-methoxy-benzenesulfonamide 814-Methoxy-N-(4′-methoxy-4-methyl- “+++” 7.92 384 biphenyl-3-yl)-benzenesulfonamide 82 4-Methoxy-N-(4-methyl-4′-methylsulfanyl- “+”8.38 400 biphenyl-3-yl)-benzenesulfonamide 834-Methoxy-N-(4-methyl-4′-trifluoromethoxy- “+” 8.74 438biphenyl-3-yl)-benzenesulfonamide 84N-(4′-Ethynyl-4-methyl-biphenyl-3-yl)- “++” 8.48 3624-methyl-benzenesulfonamide 85 N-(4′-Cyano-4-methyl-biphenyl-3-yl)-“+++” 7.67 397 3-fluoro-4-methoxy-benzenesulfonamide 86N-(4′-Cyano-4-methyl-biphenyl-3-yl)- “+” 7.67 392 (M − 1)4-nitro-benzenesulfonamide 87 4-Amino-N-(4′-cyano-4-methyl-biphenyl-“+++” 6.85 364 3-yl)-benzenesulfonamide 88N-(4′-Cyano-4-methyl-biphenyl-3-yl)- “+++” 8.04 3813-fluoro-4-methyl-benzenesulfonamide 89N-(4′-Cyano-3′-methoxy-4-methyl-biphenyl- “+++” 7.56 4093-yl)-4-methoxy-benzenesulfonamide 90N-(4′-Cyano-3′-hydroxy-4-methyl-biphenyl- “+++” 6.89 3953-yl)-4-methoxy-benzenesulfonamide 91N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “++” 7.78 373phenyl]-4-fluoro-benzenesulfonamide 92N-[5-(5-Cyano-thiophen-2-yl)-2-methyl- “+++” 7.71 403phenyl]-3-fluoro-4-methoxy- benzenesulfonamide 934-Amino-N-[5-(5-cyano-thiophen-2-yl)- “+++” 6.86 3702-methyl-phenyl]-benzenesulfonamide 944-Methyl-N-[3-methyl-5-(5-methyl-thiophen- “+++” 8.56 3583-yl)-phenyl]-benzenesulfonamide 95 N-(5-(5-cyanothiophen-3-yl)-2- “+++”8.09 385 (M − 1) methylphenyl)-3-fluoro-4- methylbenzenesulfonamide 96N-(5-(5-cyanothiophen-3-yl)-2- “+” 7.77 398 (M − 1)methylphenyl)-4-nitrobenzenesulfonamide 974-chloro-N-(4′-cyano-4-methylbiphenyl-3- “+++” 8.10 381 (M − 1)yl)benzenesulfonamide 98 N-(5-(5-cyanothiophen-2-yl)-2- “++” 6.83 369 (M− 1) methylphenyl)-4- hydroxybenzenesulfonamide 99N-(4′-cyano-3′-methoxy-4-methylbiphenyl-3- “+++” 7.99 409 (M − 1)yl)-3-fluoro-4-methylbenzenesulfonamide 100N-(4′-cyano-4-methylbiphenyl-3-yl)-4- “+” 6.38 363 (M − 1)hydroxybenzenesulfonamide 101 N-(4′-cyano-3′-methoxy-4-methylbiphenyl-3-“+” 7.63 422 (M − 1) yl)-4-nitrobenzenesulfonamide 1024-amino-N-(4′-cyano-3′-methoxy-4- “+++” 7.58 392 (M − 1)methylbiphenyl-3-yl)benzenesulfonamide 103N-(4′-cyano-3′-methoxy-4-methylbiphenyl-3- “+++” 8.46* 425 (M − 1)yl)-3-fluoro-4-methoxybenzenesulfonamide 104N-(4′-cyano-4-methylbiphenyl-3-yl)-3- “+” 8.55* 365 (M − 1)fluorobenzenesulfonamide 105 N-(4′-cyano-4-methylbiphenyl-3-yl)-3- “+”8.69* 410 (M − 1) fluoro-4-nitrobenzenesulfonamide 106N-(5-(5-aminopyridin-2-yl)-2- “+” 6.24* 352 (M − 1)methylphenyl)-4-methylbenzenesulfonamide 107N-(5-(2-methoxypyrimidin-4-yl)-2- “+” 8.43* 368 (M − 1)methylphenyl)-4-methylbenzenesulfonamide 108N-(5-(5-cyanothiophen-2-yl)-2- “++” 8.15* 415 methylphenyl)-3.4-dimethoxybenzenesulfonamide 109 N-(5-(5-cyanothiophen-2-yl)-2- “+++”8.79* 399 methylphenyl)-4-methoxy-3- methylbenzenesulfonamide 1104-amino-N-(4′-cyano-4-methylbiphenyl-3- “+++” 7.77* 380 (M − 1)yl)-3-fluorobenzenesulfonamide 111 4-methyl-N-(2-methyl-5-(6-methyl-5-“+++” 8.96* 398 nitropyridin-2-yl)phenyl)benzenesulfonamide 112N-(5-(5-cyanothiophen-2-yl)-2- “+++” 8.96* 387 methylphenyl)-4-fluoro-3-methylbenzenesulfonamide 113 N-(5-(5-cyanothiophen-2-yl)-2- “++” 8.68*401 (M − 1) methylphenyl)-4-fluoro-3- methoxybenzenesulfonamide 114N-(5-(5-cyanothiophen-2-yl)-2- “+++” 8.44* 399methylphenyl)benzo[d][1,3]dioxole-5- sulfonamide 115N-(4′-cyano-4-methylbiphenyl-3-yl)-4- “+++” 8.60* 392(dimethylamino)benzenesulfonamide 1164-cyano-N-(5-(5-cyanothiophen-2-yl)-2- “++” 8.36* 378 (M − 1)methylphenyl)benzenesulfonamide Reference: EC₅₀ <500 nM: +++ 500-1000nM: ++ 1-10 μM: + Retention times marked with “*” were measured using analternative method to the one given in Example 2. These retention timeswere determined as follows: Measured via HPLC/MS, using a WatersX-bridge C₁₈-column, 5 μm particle size, 4.6 × 150 mm (diameter ×length) at a flow rate of 1.75 ml/min with a linear gradient (water tomethanol, 0.2% formic acid as modifier) from initially 99:1 to 1:99 over9.10 min, then hold for 1.80 min. Mass signals were determined using aWaters 3100 Mass Detector.

TABLE 2B HG1 cells Example no. EC₅₀ 8 +++ 15 +++ 19 +++ 22 +++ 47 +++ 51+++ 73 +++

Example 4 Inhibition of Estrogen Receptor (ER) Signaling

To determine the effect of the compounds on estrogen receptor(ER)-mediated transcriptional activity a transfection assay using theMCF-7 cell line was performed. A luciferase-coupled ERE-tk-luc constructwas obtained from Dr. G. Reid (EMBL).

In brief, MCF-7 cells were maintained as described previously and seededon the first experimental day at a concentration of 3500 cells/well in100 μl medium and incubated under standard conditions for 24 h.Following this initial incubation period, transfection was carried outusing 5 ng of the ERE-tk-luc construct (per well) and Exgene 500transfection reagent (Fermentas) in a final buffered solution containing150 mM NaCl and 20 mM Tris pH 8.4. The plates were then maintained for3-4 hours until cell culture conditions in the incubator beforecompounds were added at 11× concentrations to yield the final desiredcompound concentrations in 0.5% DMSO (in the same manner as for theproliferation assays described previously).

After an additional 24 h incubation period the degree of inhibition ofER-signaling was determined via a luciferase readout. Cells were treatedwith Britelite™ plus solution according to the manufacturer'sinstructions (PerkinElmer, britelite plus, Ultra-High SensitivityLuminescence Reporter Gene Assay System) and luciferase levels measuredon an Envision HTS multilabel plate reader (PerkinElmer) in luminescencemode according to established protocols. Raw data were imported into anActivityBase database (IDBS, ID Business Solutions) and IC₅₀ valuescalculated using the IDBS program ActivityBase XE.

The results are depicted in Table 3. As shown in these experiments, thecompounds of the invention are capable of inhibiting estrogen receptorelement-mediated transcriptional activity in an ERα-positive cell line.The utility of ER-modulating agents for the treatment of breast, uterineor prostate cancer, as well as metastatic bone disease, is known fromthe literature (see for example, Park & Jordan (2002) Trends Mol. Med.8(2): 82-88; Steiner et al. (2001) Urology 57(4 Suppl 1): 68-72 andCampisi et al. (1993) Eur. J. Gynaecol. Oncol. 14(6): 479-483.)).

TABLE 3 Example no. IC₅₀  4 +++  5 +++ 15 +++ 43 +++ 47 +++ 49 +++ 60+++ 72 +++ Reference: IC₅₀ <500 nM: +++ 500-1000 nM: ++ 1-10 μM: +

Example 5 Inhibition of In Vitro Tubulin Polymerization

To assess the degree of inhibition of in vitro tubulin polymerization, atubulin solution (prepared in-house from porcine brain using a standardprocedure involving cycles of polymerization/de-polymerization, seeCastoldi & Popov (2003) Protein Expr. Purif. 32(1): 83-88) was incubatedwith varying concentrations of compounds under polymerizing conditionsand kinetic experiments performed to determine the amount ofpolymerization (as measured by changes in OD at 340 nm) taking placeover a 90 min period.

Compound solutions were diluted in double-distilled water to yield final10× solutions containing 5% DMSO. 5 μl solutions were then added to thewells on a 384-well clear-bottom plate (Corning #3711, Corning Inc.) inpreparation for the addition of tubulin. The tubulin solution (19.6mg/ml, in 80 mM K-PIPES [pH 6.8], 1 mM MgCl₂, 1 mM EGTA) wasre-suspended to a final concentration of 4 mg/ml with ice cold G-PEMbuffer (80 mM PIPES [pH 6.8], 2 mM MgCl₂, 0.5 mM EGTA, 10% glycerol, 1mM GTP) and maintained on ice for at least 1 min. 50 μl was then addedto the pre-dispensed compound solutions, the plate orbital shaken on amedium setting for 5 s, and the first measurement begun immediately.

Experiments were carried out on a Safire²™ monochromator (Tecan) presetto a stable temperature of 37° C.; the degree of polymerization wasdetermined by measuring the absorbance of the solutions at 340 nm everyminute over the 90 min cycle. Reported inhibition values were based onthe final absorbance at 90 min and calculated with reference to avehicle control (0.5% DMSO) using the minimum and maximum signalsobtained for this sample.

As demonstrated by these experiments (see Table 4), changes in thestructural properties of the various examples can lead to a selection ofcompounds that are active in inhibiting cell proliferation and affectHIF-signaling, but that do, or do not, have tubulin inhibitionproperties, depending on whether this is desired in the target productprofile.

TABLE 4 Example no. Inhibition (%)  3 + 11 +/− 16 +/− 23 + 56 + 60 − 63− 72 − Reference: Inhibition at 10 μM >50%: + 20-50%: +/− <20% −

Example 6 Inhibition of Activation of HIF Mediated Transcription underHypoxic Conditions

Inhibition of an activated HIF signaling response underchemically-induced hypoxic conditions due to compound treatment wasdetermined using the CellSensor® HRE-bla HCT-116 stably transfectedreporter cell line from Invitrogen according to the manufacturer'sinstructions. HIF is a transcriptional factor composed of theconstitutively expressed HIF1β subunit and one of three HIFα subunits(HIF1α, HIF2α, HIF3α). This assay is responsive towards HIFa activity ingeneral. In one example, HIF-1, when stabilized by hypoxic conditions,upregulates several genes to promote cell survival in low-oxygenconditions. These include glycolysis enzymes, which allow ATP synthesisin an oxygen-independent manner, and vascular endothelial growth factor(VEGF), which promotes angiogenesis. HIF-1 acts by binding toHIF-responsive elements (HREs) in promoters that generally contain thesequence NCGTG.

Cells were maintained as described previously and seeded into 384-well,clear-bottom plates (Corning 3712) at 15000 cells/well in 32 μl assaymedium (Opti-MEM [Invitrogen], 0.5% FBS, 100 U/ml penicillin, 100 μg/mlstreptomycin, 0.1 mM non-essential amino acids [NEAA], 1 mM sodiumpyruvate, 5 mM HEPES [pH 7.3]). Following a 2 h incubation period,compounds (4 μl) were subsequently added to the cells at 10×concentrations in 5% DMSO and incubated under normal conditions for 30min. To induce hypoxic conditions, 4 μl of a 2 mM deferoxamine (DFO)solution was added to the cells followed by 24 h incubation understandard assay conditions (as described). Control wells included wellscontaining only medium (no cells) and wells treated with 0.5% DMSOinstead of compound.

Prior to the readout, the Substrate Loading Solution was prepared asdescribed in the manufacturer's protocol and 10 μl added to each well.Following a further 2 h incubation period at room temperature and in thedark, fluorescence was measured at two wavelengths (blue channel: ex.409 nm, em. 460 nm; green channel: ex. 409 nm, em. 530 nm) on aPerkinElmer Envision HTS. For the analysis, the average signal of thecell-free wells at 460 nm and 530 nm was first subtracted from the blueand green channel data, respectively. The blue/green emission ratioswere then calculated for each well, dividing the background-correctedblue emission values by the background-corrected green emission values.IC₅₀ values were determined from these ratios using GraphPad Prism(Prism 5, GraphPad software, Inc.).

The results of these experiments (see Table 5) show that the compoundsof the invention are capable of inhibiting hypoxia regulatedelement-mediated transcriptional activity under hypoxic conditions. Thecompounds of this invention have a potency level that exceedsHIF-inhibitor compounds described in the prior art such as, e.g. theProlX compound PX-478, the only small molecule HIF-inhibitor currentlyreported to undergo clinical trials.

TABLE 5 Example no. EC₅₀  3 +++  8 +++ 23 +++ 43 +++ 51 +++ 68 +++ 71+++ 73 +++ 95 +++ 103  +++ 112  +++ 114  +++ Reference: EC₅₀ <500 nM:+++ 500-1000 nM: ++ 1-10 μM: +

Example 7 Activation of Apoptosis

Activation of caspase-3/7 signal following treatment with test compoundswas carried out using a Caspase-Glo® 3/7 kit (Promega). In brief, HG-1cells were seeded at a concentration of 1000 cells/well in 50 μl mediuminto 96-well plates and incubated for 24 h under the conditionsdescribed previously. Compounds at various concentrations were thenadded to the wells (at all final concentrations 0.5% DMSO) and incubatedfor a further 24 h period. The plates were subsequently removed from theincubator and allowed to equilibrate to room temperature, after which 50μl Caspase-Glo reagent was added to all wells, the plate shaken at 300rpm for 30 s, and luminescence measured after a 30 min incubation periodon an Envision HTS (PerkinElmer).

Following treatment with compounds of the invention, a clear activationof the caspase-3/7 response was observed (see Table 6), indicating thatapoptosis is one factor in the observed decrease in cell proliferationdue to the compound mechanisms of action.

TABLE 6 Example no. EC₅₀ 47 +++ 51 +++ 56 +++ 60 +++ 63 +++ Reference:EC₅₀ <500 nM: +++ 500-1000 nM: ++ 1-10 μM: +

Example 8 Cell Cycle Analysis

To determine the effect on the cell cycle of HL-60 cells followingtreatment with compounds, fluorescence-assisted cell sorting (FACS) wasused, sorting cells by nuclear staining.

A standard protocol using propidium iodide was utilized. In brief, cellswere seeded into 6-well tissue culture plates at 2×10⁵ cells in 2 mlmedium and incubated for 24 h at 37° C. under standard tissue cultureconditions (humidified 95% air, 5% CO₂). Following this initialincubation period, compound solutions were added from stock solutionsyielding final concentrations of 20 μM in 0.1% DMSO and the cellsincubated after gentle mixing for further 24 h.

Cells were subsequently harvested, washed, and re-suspended inphosphate-bufferd saline (PBS, pH 7.4). Fixation was performed inice-cold 70% ethanol while vortexing at half-speed and the solutionsstored overnight at 4° C. The cells were then spun down, washed 3 timesin PBS, and stained with a propidium iodide staining solution (20 μg/mlpropidium iodide, 200 μg/ml RNase A, 0.1% Triton X-100 in PBS, pH 7.4).Finally, samples were passed through a 70 μm filter (BD Falcon) prior tomeasurement on a FACScan™ flow cytometer (Beckton Dickinson).

Together with the data showing caspase-3/7 activation, the FACS analysisshown for a representative compound demonstrates the appearance of asub-2N population indicative of a cell cycle effect leading to apoptosis(see results depicted in FIG. 1). The ability of the compounds of theinvention to induce apoptosis is a useful feature which can be exploitedin chemotherapeutical applications.

Example 9 Further Useful Compounds According to the Invention

Further compounds of the invention are exemplified in Table 7, which areof particular use within the scope of the present invention. Thesecompounds can be produced as outlined under example 1 and also accordingto manufacturing practice well known in organic chemistry as describedabove. These compounds will have a similar inhibitory activity as shownfor the compounds listed in table 2A and tested in examples 3 to 8.

TABLE 7 Example Number IUPAC Name 1174-Methyl-N-[3-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide118 N-(3-Furan-2-yl-5-methyl-phenyl)-4-methyl-benzenesulfonamide 119N-(3-Furan-3-yl-5-methyl-phenyl)-4-methyl-benzenesulfonamide 1204-Methyl-N-(3-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide 1214-Methyl-N-[3-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide122 4-Methyl-N-(3-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide 123N-[3-(2,4-Dimethyl-thiazol-5-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide124 4-Methyl-N-(3-methyl-5-pyrrol-1-yl-phenyl)-benzenesulfonamide 125N-[3-(3,5-Dimethyl-isoxazol-4-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide1264-Methyl-N-[3-methyl-5-(2-methyl-thiophen-3-yl)-phenyl]-benzenesulfonamide127 4-Methyl-N-(3-methyl-5-oxazol-5-yl-phenyl)-benzenesulfonamide 1284-Methyl-N-[3-methyl-5-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide129N-[3-(6-Cyano-pyridin-3-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide130N-[3-(5-Cyano-pyridin-2-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide131N-[3-(5-Methoxy-pyridin-2-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide132N-[3-(5-Cyano-thiophen-2-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide133N-[3-(5-Fluoro-pyridin-2-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide134 N-(4′-Cyano-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide 135N-(3′,4′-Dicyano-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide 136N-(4′-Cyano-3′-fluoro-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide137 N-(4′-Cyano-5,3′-dimethyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide138N-[3-(2-Cyano-pyridin-4-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide1394-Methyl-N-[3-methyl-5-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide1404-Chloro-N-[3-(5-cyano-thiophen-2-yl)-5-methyl-phenyl]-benzenesulfonamide141 N-(4′-Methoxy-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide142N-(4′-Cyano-3′-hydroxy-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide143N-(4′-Cyano-3′-methoxy-5-methyl-biphenyl-3-yl)-4-methyl-benzenesulfonamide144N-(4′-Cyano-3′-hydroxy-5-methyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide145N-(4′-Cyano-3′-methoxy-5-methyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide146N-[3-(5-Cyano-pyridin-2-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide1474-Methoxy-N-[3-(5-methoxy-pyridin-2-yl)-5-methyl-phenyl]-benzenesulfonamide148N-[3-(5-Cyano-thiophen-2-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide149N-[3-(5-Fluoro-pyridin-2-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide150 N-(4′-Cyano-5-methyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide 151N-(3′,4′-Dicyano-5-methyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide152N-(4′-Cyano-3′-fluoro-5-methyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide153N-(4′-Cyano-5,3′-dimethyl-biphenyl-3-yl)-4-methoxy-benzenesulfonamide154N-[3-(2-Cyano-pyridin-4-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide1554-Methoxy-N-[3-methyl-5-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide156 4-Methoxy-N-(4′-methoxy-5-methyl-biphenyl-3-yl)-benzenesulfonamide157 4-Methoxy-N-(3-methyl-5-pyrrol-1-yl-phenyl)-benzenesulfonamide 158N-[3-(3,5-Dimethyl-isoxazol-4-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide1594-Methoxy-N-[3-methyl-5-(2-methyl-thiophen-3-yl)-phenyl]-benzenesulfonamide160 4-Methoxy-N-(3-methyl-5-oxazol-5-yl-phenyl)-benzenesulfonamide 1614-Methoxy-N-[3-methyl-5-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide162N-[3-(6-Cyano-pyridin-3-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide1634-Methoxy-N-[3-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide164 N-(3-Furan-2-yl-5-methyl-phenyl)-4-methoxy-benzenesulfonamide 165N-(3-Furan-3-yl-5-methyl-phenyl)-4-methoxy-benzenesulfonamide 1664-Methoxy-N-(3-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide 1674-Methoxy-N-[3-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide168 4-Methoxy-N-(3-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide 169N-[3-(2,4-Dimethyl-thiazol-5-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide170N-[3-(2,4-Dimethyl-thiazol-5-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 1714-Methoxy-N-methyl-N-(4-methyl-3-thiophen-2-yl-phenyl)-benzenesulfonamide1724,N-Dimethyl-N-[4-methyl-3-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide173 4,N-Dimethyl-N-(4-methyl-3-thiophen-3-yl-phenyl)-benzenesulfonamide174 4-Methoxy-N-methyl-N-[4-methyl-3-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide 1754-Methoxy-N-methyl-N-(4-methyl-3-thiophen-3-yl-phenyl)-benzenesulfonamide176N-(3-Furan-3-yl-4-methyl-phenyl)-4-methoxy-N-methyl-benzenesulfonamide177 N-[3-(6-Cyano-pyridin-3-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 1784-Methoxy-N-methyl-N-(4-methyl-3-oxazol-5-yl-phenyl)-benzenesulfonamide179 4-Methoxy-N-methyl-N-[4-methyl-3-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide 1804-Methoxy-N-methyl-N-[4-methyl-3-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 181N-(3-Furan-2-yl-4-methyl-phenyl)-4-methoxy-N-methyl-benzenesulfonamide182 4-Methoxy-N-methyl-N-[4-methyl-3-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide 1834-Methoxy-N-methyl-N-[4-methyl-3-(2-methyl-thiophen-3-yl)-phenyl]-benzenesulfonamide 184N-[3-(3,5-Dimethyl-isoxazol-4-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 185N-[3-(2-Cyano-pyridin-4-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 1864-Methoxy-N-(4′-methoxy-6-methyl-biphenyl-3-yl)-N-methyl-benzenesulfonamide1874-Methoxy-N-methyl-N-(4-methyl-3-pyrrol-1-yl-phenyl)-benzenesulfonamide188N-(4′-Cyano-6,3′-dimethyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide189N-(4′-Cyano-6-methyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide190N-(3′,4′-Dicyano-6-methyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide191 N-(4′-Cyano-3′-fluoro-6-methyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide 1924,N-Dimethyl-N-(4-methyl-3-thiophen-2-yl-phenyl)-benzenesulfonamide 1934-Methoxy-N-[3-(5-methoxy-pyridin-2-yl)-4-methyl-phenyl]-N-methyl-benzenesulfonamide 194N-[3-(5-Cyano-thiophen-2-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 195N-[3-(5-Fluoro-pyridin-2-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide196 N-(4′-Cyano-3′-hydroxy-6-methyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide 197N-[3-(5-Cyano-pyridin-2-yl)-4-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 198N-(4′-Cyano-3′-methoxy-6-methyl-biphenyl-3-yl)-4-methoxy-N-methyl-benzenesulfonamide 199N-(4′-Cyano-3′-methoxy-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide200 N-(3-Furan-3-yl-4-methyl-phenyl)-4,N-dimethyl-benzenesulfonamide 2014,N-Dimethyl-N-[4-methyl-3-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide202 N-[3-(3,5-Dimethyl-isoxazol-4-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide 203N-[3-(2,4-Dimethyl-thiazol-5-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide 204N-[3-(5-Methoxy-pyridin-2-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide205 4,N-Dimethyl-N-(4-methyl-3-pyrrol-1-yl-phenyl)-benzenesulfonamide206 4,N-Dimethyl-N-(4-methyl-3-oxazol-5-yl-phenyl)-benzenesulfonamide2074,N-Dimethyl-N-[4-methyl-3-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide208N-[3-(5-Cyano-pyridin-2-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide2094,N-Dimethyl-N-[4-methyl-3-(2-methyl-thiophen-3-yl)-phenyl]-benzenesulfonamide210N-[3-(5-Cyano-thiophen-2-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide211 N-(3-Furan-2-yl-4-methyl-phenyl)-4,N-dimethyl-benzenesulfonamide 212N-[3-(6-Cyano-pyridin-3-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide213 N-(4′-Cyano-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide214N-(3′,4′-Dicyano-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide215N-(4′-Cyano-3′-hydroxy-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide216N-[3-(2-Cyano-pyridin-4-yl)-4-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide217N-(4′-Cyano-3′-fluoro-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide218N-(4′-Cyano-6,3′-dimethyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide2194,N-Dimethyl-N-[4-methyl-3-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide220N-(4′-Methoxy-6-methyl-biphenyl-3-yl)-4,N-dimethyl-benzenesulfonamide221 4-Chloro-N-[3-(5-cyano-thiophen-2-yl)-4-methyl-phenyl]-N-methyl-benzenesulfonamide 2224-Methyl-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide2234-Methoxy-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide2244-Methyl-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide2254-Methoxy-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide2264-Methyl-N-[3-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide227 4-Methoxy-N-methyl-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide 2284-Methyl-N-[3-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide229 4-Methoxy-N-methyl-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide 2304,N-Dimethyl-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide 2314-Methoxy-N-methyl-N-[3-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide 2324,N-Dimethyl-N-[3-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide 2334-Methoxy-N-[3-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide2344-Amino-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide2354-Hydroxy-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide2364-Chloro-N-[2-methyl-5-(1-methyl-1H-pyrrol-2-yl)-phenyl]-benzenesulfonamide2374-Chloro-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide2384-Hydroxy-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide2394-Amino-N-[2-methyl-5-(1-methyl-1H-pyrrol-3-yl)-phenyl]-benzenesulfonamide240N-[5-(1,3-Dimethyl-1H-pyrrol-2-yl)-2-methyl-phenyl]-4-methyl-benzenesulfonamide241 N-[5-(1,3-Dimethyl-1H-pyrrol-2-yl)-2-methyl-phenyl]-4-methoxy-benzenesulfonamide 242N-[5-(1,4-Dimethyl-1H-pyrrol-3-yl)-2-methyl-phenyl]-4-methyl-benzenesulfonamide243 N-[5-(1,4-Dimethyl-1H-pyrrol-3-yl)-2-methyl-phenyl]-4-methoxy-benzenesulfonamide 244N-[3-(1,3-Dimethyl-1H-pyrrol-2-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide245N-[5-(1,3-Dimethyl-1H-pyrrol-2-yl)-2-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 246N-[3-(1,4-Dimethyl-1H-pyrrol-3-yl)-5-methyl-phenyl]-4-methyl-benzenesulfonamide247N-[5-(1,4-Dimethyl-1H-pyrrol-3-yl)-2-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 248N-[5-(1,3-Dimethyl-1H-pyrrol-2-yl)-2-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide 249N-[3-(1,3-Dimethyl-1H-pyrrol-2-yl)-5-methyl-phenyl]-4-methoxy-N-methyl-benzenesulfonamide 250N-[3-(1,4-Dimethyl-1H-pyrrol-3-yl)-5-methyl-phenyl]-4,N-dimethyl-benzenesulfonamide 251N-[3-(1,4-Dimethyl-1H-pyrrol-3-yl)-5-methyl-phenyl]-4-methoxy-benzenesulfonamide 2524-Hydroxy-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide2534-Amino-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide2544-Chloro-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide2554-Hydroxy-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide2564-Amino-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide2574-Chloro-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide258 N-(5-Furan-2-yl-2-methyl-phenyl)-4-hydroxy-benzenesulfonamide 2594-Amino-N-(5-furan-2-yl-2-methyl-phenyl)-benzenesulfonamide 2604-Chloro-N-(5-furan-2-yl-2-methyl-phenyl)-benzenesulfonamide 2614-Hydroxy-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide 2624-Amino-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide 2634-Chloro-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide 2644-Hydroxy-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide 2654-Amino-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide 2664-Chloro-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide 267N-(5-Furan-3-yl-2-methyl-phenyl)-4-hydroxy-benzenesulfonamide 2684-Amino-N-(5-furan-3-yl-2-methyl-phenyl)-benzenesulfonamide 2694-Chloro-N-(5-furan-3-yl-2-methyl-phenyl)-benzenesulfonamide 2704-Hydroxy-N-methyl-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 2714-Amino-N-methyl-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 2724-Chloro-N-methyl-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 2734-Hydroxy-N-methyl-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide 2744-Amino-N-methyl-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide 2754-Chloro-N-methyl-N-[2-methyl-5-(5-methyl-thiophen-2-yl)-phenyl]-benzenesulfonamide 276N-(5-Furan-2-yl-2-methyl-phenyl)-4-hydroxy-N-methyl-benzenesulfonamide277 4-Amino-N-(5-furan-2-yl-2-methyl-phenyl)-N-methyl-benzenesulfonamide2784-Chloro-N-(5-furan-2-yl-2-methyl-phenyl)-N-methyl-benzenesulfonamide2794-Hydroxy-N-methyl-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide2804-Amino-N-methyl-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide2814-Chloro-N-methyl-N-(2-methyl-5-thiophen-2-yl-phenyl)-benzenesulfonamide2824-Hydroxy-N-methyl-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide2834-Amino-N-methyl-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide2844-Chloro-N-methyl-N-(2-methyl-5-thiophen-3-yl-phenyl)-benzenesulfonamide285N-(5-Furan-3-yl-2-methyl-phenyl)-4-hydroxy-N-methyl-benzenesulfonamide286 4-Amino-N-(5-furan-3-yl-2-methyl-phenyl)-N-methyl-benzenesulfonamide2874-Chloro-N-(5-furan-3-yl-2-methyl-phenyl)-N-methyl-benzenesulfonamide288 N-[5-(5-Cyano-furan-2-yl)-2-methyl-phenyl]-4-hydroxy-N-methyl-benzenesulfonamide 2894-Amino-N-[5-(5-cyano-furan-2-yl)-2-methyl-phenyl]-N-methyl-benzenesulfonamide 2904-Chloro-N-[5-(5-cyano-furan-2-yl)-2-methyl-phenyl]-N-methyl-benzenesulfonamide 2914-Hydroxy-N-methyl-N-[2-methyl-5-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide 2924-Amino-N-methyl-N-[2-methyl-5-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide2934-Chloro-N-methyl-N-[2-methyl-5-(5-nitro-furan-2-yl)-phenyl]-benzenesulfonamide294N-[5-(5-Cyano-thiophen-3-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide2954-Amino-N-[5-(5-cyano-thiophen-3-yl)-2-methyl-phenyl]-benzenesulfonamide2964-Chloro-N-[5-(5-cyano-thiophen-3-yl)-2-methyl-phenyl]-benzenesulfonamide297N-[5-(5-Cyano-furan-2-yl)-2-methyl-phenyl]-N-methyl-4-trifluoromethoxy-benzenesulfonamide 298N-[5-(5-Cyano-furan-2-yl)-2-methyl-phenyl]-N-methyl-4-trifluoromethyl-benzenesulfonamide 299N-[5-(5-Cyano-thiophen-3-yl)-2-methyl-phenyl]-4-trifluoromethoxy-benzenesulfonamide 300N-[5-(5-Cyano-thiophen-3-yl)-2-methyl-phenyl]-4-trifluoromethyl-benzenesulfonamide 301N-[5-(5-Cyano-furan-2-yl)-2-methyl-phenyl]-4-trifluoromethoxy-benzenesulfonamide 302N-[5-(5-Cyano-furan-2-yl)-2-methyl-phenyl]-4-trifluoromethyl-benzenesulfonamide303N-[5-(2,4-Dimethyl-thiazol-5-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide304N-[5-(3,5-Dimethyl-isoxazol-4-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide3054-Amino-N-[5-(2,4-dimethyl-thiazol-5-yl)-2-methyl-phenyl]-benzenesulfonamide3064-Chloro-N-[5-(2,4-dimethyl-thiazol-5-yl)-2-methyl-phenyl]-benzenesulfonamide3074-Amino-N-[5-(3,5-dimethyl-isoxazol-4-yl)-2-methyl-phenyl]-benzenesulfonamide3084-Chloro-N-[5-(3,5-dimethyl-isoxazol-4-yl)-2-methyl-phenyl]-benzenesulfonamide309 4-Hydroxy-N-(2-methyl-5-oxazol-5-yl-phenyl)-benzenesulfonamide 3104-Amino-N-(2-methyl-5-oxazol-5-yl-phenyl)-benzenesulfonamide 3114-Chloro-N-(2-methyl-5-oxazol-5-yl-phenyl)-benzenesulfonamide 3124-Hydroxy-N-(2-methyl-5-pyrrol-1-yl-phenyl)-benzenesulfonamide 3134-Amino-N-(2-methyl-5-pyrrol-1-yl-phenyl)-benzenesulfonamide 3144-Chloro-N-(2-methyl-5-pyrrol-1-yl-phenyl)-benzenesulfonamide 3154-Hydroxy-N-[2-methyl-5-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide3164-Amino-N-[2-methyl-5-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide3174-Chloro-N-[2-methyl-5-(3-methyl-pyrazol-1-yl)-phenyl]-benzenesulfonamide3184-Hydroxy-N-[2-methyl-5-(2-methyl-pyrrol-1-yl)-phenyl]-benzenesulfonamide3194-Amino-N-[2-methyl-5-(2-methyl-pyrrol-1-yl)-phenyl]-benzenesulfonamide3204-Chloro-N-[2-methyl-5-(2-methyl-pyrrol-1-yl)-phenyl]-benzenesulfonamide321 N-(4,2′-Dimethyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide 3224-Amino-N-(4,2′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 3234-Chloro-N-(4,2′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 324N-(4,3′-Dimethyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide 3254-Amino-N-(4,3′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 3264-Chloro-N-(4,3′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 327N-(4,4′-Dimethyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide 3284-Amino-N-(4,4′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 3294-Chloro-N-(4,4′-dimethyl-biphenyl-3-yl)-benzenesulfonamide 3304-Hydroxy-N-(2-methyl-5-pyridin-2-yl-phenyl)-benzenesulfonamide 3314-Amino-N-(2-methyl-5-pyridin-2-yl-phenyl)-benzenesulfonamide 3324-Chloro-N-(2-methyl-5-pyridin-2-yl-phenyl)-benzenesulfonamide 3334-Hydroxy-N-(2-methyl-5-pyridin-3-yl-phenyl)-benzenesulfonamide 3344-Amino-N-(2-methyl-5-pyridin-3-yl-phenyl)-benzenesulfonamide 3354-Chloro-N-(2-methyl-5-pyridin-3-yl-phenyl)-benzenesulfonamide 3364-Hydroxy-N-(2-methyl-5-pyridin-4-yl-phenyl)-benzenesulfonamide 3374-Amino-N-(2-methyl-5-pyridin-4-yl-phenyl)-benzenesulfonamide 3384-Chloro-N-(2-methyl-5-pyridin-4-yl-phenyl)-benzenesulfonamide 339N-[5-(5-Cyano-pyridin-3-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide3404-Amino-N-[5-(5-cyano-pyridin-3-yl)-2-methyl-phenyl]-benzenesulfonamide3414-Chloro-N-[5-(5-cyano-pyridin-3-yl)-2-methyl-phenyl]-benzenesulfonamide3424-Hydroxy-N-[2-methyl-5-(6-methyl-pyridin-2-yl)-phenyl]-benzenesulfonamide3434-Amino-N-[2-methyl-5-(6-methyl-pyridin-2-yl)-phenyl]-benzenesulfonamide3444-Chloro-N-[2-methyl-5-(6-methyl-pyridin-2-yl)-phenyl]-benzenesulfonamide345N-[5-(5-Cyano-pyridin-2-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide3464-Amino-N-[5-(5-cyano-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide3474-Chloro-N-[5-(5-cyano-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide348 N-[5-(5-Cyano-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethoxy-benzenesulfonamide 349N-[5-(5-Cyano-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethyl-benzenesulfonamide 3504-Hydroxy-N-[5-(5-methoxy-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide3514-Amino-N-[5-(5-methoxy-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide3524-Chloro-N-[5-(5-methoxy-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide353 N-[5-(5-Methoxy-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethoxy-benzenesulfonamide 354N-[5-(5-Methoxy-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethyl-benzenesulfonamide 355N-[5-(5-Fluoro-pyridin-2-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide3564-Amino-N-[5-(5-fluoro-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide3574-Chloro-N-[5-(5-fluoro-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide358 N-[5-(5-Fluoro-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethoxy-benzenesulfonamide 359N-[5-(5-Fluoro-pyridin-2-yl)-2-methyl-phenyl]-4-trifluoromethyl-benzenesulfonamide 3604-Hydroxy-N-[2-methyl-5-(5-nitro-pyridin-2-yl)-phenyl]-benzenesulfonamide3614-Amino-N-[2-methyl-5-(5-nitro-pyridin-2-yl)-phenyl]-benzenesulfonamide3624-Chloro-N-[2-methyl-5-(5-nitro-pyridin-2-yl)-phenyl]-benzenesulfonamide363 N-[2-Methyl-5-(5-nitro-pyridin-2-yl)-phenyl]-4-trifluoromethoxy-benzenesulfonamide 364N-[2-Methyl-5-(5-nitro-pyridin-2-yl)-phenyl]-4-trifluoromethyl-benzenesulfonamide365 4-Amino-N-(4′-cyano-4-methyl-biphenyl-3-yl)-benzenesulfonamide 366N-[5-(6-Cyano-pyridin-3-yl)-2-methyl-phenyl]-4-hydroxy-benzenesulfonamide3674-Amino-N-[5-(6-cyano-pyridin-3-yl)-2-methyl-phenyl]-benzenesulfonamide3684-Chloro-N-[5-(6-cyano-pyridin-3-yl)-2-methyl-phenyl]-benzenesulfonamide369N-(3′,4′-Dicyano-4-methyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide370 4-Amino-N-(3′,4′-dicyano-4-methyl-biphenyl-3-yl)-benzenesulfonamide371 4-Chloro-N-(3′,4′-dicyano-4-methyl-biphenyl-3-yl)-benzenesulfonamide372N-(4′-Cyano-3′-fluoro-4-methyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide3734-Amino-N-(4′-cyano-3′-fluoro-4-methyl-biphenyl-3-yl)-benzenesulfonamide3744-Chloro-N-(4′-cyano-3′-fluoro-4-methyl-biphenyl-3-yl)-benzenesulfonamide375N-(4′-Cyano-4,3′-dimethyl-biphenyl-3-yl)-4-hydroxy-benzenesulfonamide376 4-Amino-N-(4′-cyano-4,3′-dimethyl-biphenyl-3-yl)-benzenesulfonamide377 4-Chloro-N-(4′-cyano-4,3′-dimethyl-biphenyl-3-yl)-benzenesulfonamide3783-Fluoro-N-(5-furan-2-yl-2-methyl-phenyl)-4-methyl-benzenesulfonamide3793-Fluoro-N-(5-furan-2-yl-2-methyl-phenyl)-4-methoxy-benzenesulfonamide380 3-Fluoro-4-methyl-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 3813-Fluoro-4-methoxy-N-[2-methyl-5-(5-methyl-furan-2-yl)-phenyl]-benzenesulfonamide 382N-(4,4′-Dimethyl-biphenyl-3-yl)-3-fluoro-4-methyl-benzenesulfonamide 383N-(4,4′-Dimethyl-biphenyl-3-yl)-3-fluoro-4-methoxy-benzenesulfonamide384 3-Fluoro-N-[5-(5-methoxy-pyridin-2-yl)-2-methyl-phenyl]-4-methyl-benzenesulfonamide 3853-Fluoro-4-methoxy-N-[5-(5-methoxy-pyridin-2-yl)-2-methyl-phenyl]-benzenesulfonamide 3863-Fluoro-4-methyl-N-(2-methyl-5-pyridin-3-yl-phenyl)-benzenesulfonamide3873-Fluoro-4-methoxy-N-(2-methyl-5-pyridin-3-yl-phenyl)-benzenesulfonamide388 N-[5-(4-Cyano-thiophen-2-yl)-2-methyl-phenyl]-3-fluoro-4-methyl-benzenesulfonamide 389N-[5-(4-Cyano-thiophen-2-yl)-2-methyl-phenyl]-3-fluoro-4-methoxy-benzenesulfonamide 390N-[5-(5-Cyano-thiophen-3-yl)-2-methyl-phenyl]-3-fluoro-4-methoxy-benzenesulfonamide

Discussion

It follows directly from the data shown in the examples above that thecompounds of the invention have the common property of inhibiting HIFfunction and inhibiting cell proliferation. Thus, these compounds areuseful as therapeutic compounds for the treatment or prevention of adisease or disorder selected from the group consisting of aninflammatory disease, a hyperproliferative disease or disorder, ahypoxia-related pathology and a disease characterized by excessivevascularisation.

In addition to the above described useful properties that are shared byall compounds of the invention, these compounds also exhibit differentdegrees of potency with respect to inhibiting tubulin organization. Thisshows a further valuable contribution of this invention which lies inthe provision of therapeutic compounds that can be administeredselectively on a case to case basis to patients depending on theirdisease state, constitution and genetic predisposition. In this contextit is further possible to decide if the patient is to be treated withcompounds of the invention that do or do not possess an additionalactivity as tubulin inhibitors.

1. A compound having a structure according to formula I:

wherein R¹ is selected from a group consisting of H, alkyl, alkenyl,alkynyl, —CN, halogen, —OH, alkoxy, —SH, S-alkyl, —NH₂, NH-alkyl,N-bis-alkyl, NHOH, NMeOH, NMe(OMe), —NO₂, —CF₃, —OCF₃ and C₁-C₄hydroxyalkyl. R² is H or C₁-C₄ alkyl; R³ is H or —CH₃; R⁴ is phenyl ormonocyclic 5- or 6-membered heteroaryl; optionally substituted with oneor more substituents selected from the group consisting of: alkyl,alkenyl, alkynyl, alkoxy, halogen, —CN, —CF₃, —OCF₃, C₁-C₄ hydroxyalkyl,—OH, —SH, S-alkyl, —CN, N-bis-alkyl, cyanoacetylene, —NO₂, —NR⁷R⁸,—C(O)R²⁰, N—O (wherein the nitrogen atom is integral part of themonocyclic 5- or 6-membered heteroaryl) and two substituents which formtogether a dioxymethylene bridge (—O—CH₂—O—); R⁵ is H or —CH₃; R⁶ isselected from the group consisting of H, halogen, alkyl, alkoxy,alkenyl, alkynyl, S-alkyl, —OH, —NR⁷R⁸, —CN, N-bis-alkyl, —SH, —CF₃ and—OCF₃; or R⁶ forms together with R¹ a dioxymethylene bridge (—O—CH₂—O—);R⁷ is H or alkyl; R⁸ is H or C₁-C₄ alkyl; and R²⁰ is C₁-C₄ alkyl; withthe proviso that R⁴ is not 3-alkoxy-pyridazine-5-yl; that if R⁴ isphenyl, then the 2- and 5-position of the phenyl ring may not besubstituted with two methoxy substituents at the same time; and that R³and R⁵ are not at the same time H.
 2. The compound according to claim 1,wherein the compound has a structure according to formula II:


3. The compound according to claim 1, wherein the compound has astructure according to formula III:


4. The compound according to claim 1, wherein R⁴ has a structureaccording to formula IV:

wherein R⁹ and R¹⁰ are each individually selected from the groupconsisting of H, alkyl, C₁-C₄ alkenyl, alkynyl, —CN, —C(O)R²⁰,cyanoacetylene, halogen, —OH, C₁-C₄ alkoxy, —SH, C₁-C₄ S-alkyl, —NH₂,C₁-C₄ NH-alkyl, C₁-C₄ N-bis-alkyl, —NO₂, —CF₃, —OCF₃, and C₁-C₄hydroxyalkyl, or R⁹ and R¹⁰ form together a dioxymethylene bridge(—O—CH₂—O—); R¹¹ and R¹² are each individually selected from the groupconsisting of H, C₁-C₄ alkyl, C₁-C₄ alkenyl, C₁-C₄ alkynyl, —CN,halogen, —OH, alkoxy, —SH, C₁-C₄ S-alkyl, —CF₃, —OCF₃, —NH₂, —N(CH₃)₂and C₁-C₄ hydroxyalkyl; with the proviso that R⁹ and R¹² may not bemethoxy at the same time; R²⁰ has the above indicated meaning; and *indicates the bond between R⁴ and the compound according to any offormulas (I) through (III).
 5. The compound according to claim 4,wherein R¹¹ and R¹² are H.
 6. The compound according to claim 5, whereinR³ is methyl; R² is H, methyl or ethyl; and R⁵ and R⁶ are H.
 7. Thecompound according to claim 1, wherein R⁴ has a structure according toformula V:

wherein A, B, D and E are each individually selected from the groupconsisting of a nitrogen atom, CR¹³ and N—O; G is selected from thegroup consisting of an oxygen atom, a sulphur atom and NR¹⁴; R¹³ isselected from the group consisting of H, C₁-C₃ alkyl, C₁-C₃ alkoxy, —OH,—SH, —CF₃, —OCF₃, halogen, —NR¹⁵R¹⁶, —NO₂, —CN, —C(O)R²⁰, acetylene,cyanoacetylene, C₁-C₄ hydroxyalkyl and a σ (sigma) bond connecting R⁴ tothe compound according to any of formulas (I) through (III); and R¹⁴ isselected from the group consisting of H, C₁-C₄ alkyl and a σ (sigma)bond connecting R⁴ to the compound according to any of formulas (I)through (III); and R¹⁵ and R¹⁶ are each individually either H or C₁-C₄alkyl; R²⁰ has the above indicated meaning; and * indicates the bondbetween R⁴ and the compound according to any of formulas (I) through(III).
 8. The compound according to claim 1, wherein R⁴ has a structureaccording to formula VI:

wherein L and T are each individually either a CH group or a nitrogenatom or N—O; M, N and Q are each individually selected from the groupconsisting of a nitrogen atom, a CR¹⁷ group and N—O; R¹⁷ is selectedfrom the group consisting of H, C₁-C₃ alkyl, C₁-C₃ alkoxy, —CF₃, —OCF₃,halogen, —OH, —NO₂, —SH, C₁-C₃ S-alkyl, —NR¹⁵R¹⁶, C₁-C₄ hydroxyalkyl,—C(O)R²⁰, acetylene, cyanoacetylene and —CN; R¹⁵ and R¹⁶ are eachindividually either H or C₁-C₄ alkyl; R²⁰ has the above indicatedmeaning; and * indicates the bond between R⁴ and the compound accordingto any of formulas (I) through (III).
 9. The compound according to claim1, wherein R⁴ is selected from the group consisting of:

wherein R¹⁸ and R¹⁹ are each individually selected from the groupconsisting of H, C₁-C₃ alkyl, C₁-C₃ alkoxy, —CF₃, —OCF₃, halogen, —OH,—NO₂, —SH, C₁-C₃ S-alkyl, —NR¹⁵R¹⁶, C₁-C₄ hydroxyalkyl, alkynyl,alkenyl, —C(O)R²⁰, cyanoacetylene and —CN; and R¹⁵, R¹⁶ and R²⁰ have theabove indicated meaning.
 10. The compound according to claim 1, whereinR² is H and R⁴ is selected from the group consisting of:

wherein * indicates the bond between R⁴ and the compound according toany of formulas (I) through (III).
 11. The compound according to claim1, wherein R² and/or R⁶ is H.
 12. A compound according to claim 1 orpharmaceutically acceptable salt thereof, for the prevention ortreatment of a disease or disorder.
 13. A pharmaceutical compositioncomprising a compound according to claim 1 or pharmaceuticallyacceptable salt thereof and a second therapeutic agent useful for thetreatment or prevention of a disease or disorder selected from the groupconsisting of an inflammatory disease, a hyperproliferative disease ordisorder, a hypoxia related pathology and a disease characterized bypathophysiological hyper-vascularisation, and, optionally, apharmaceutically acceptable carrier or excipient.
 14. A method ofpreparing a medicament, comprising the steps of: providing a compoundaccording to claim 1, or a pharmaceutically acceptable salt thereof; andusing said compound for the preparation of a medicament for thetreatment or prevention of a disease or disorder selected from the groupconsisting of an inflammatory disease, a hyperproliferative disease ordisorder, a hypoxia related pathology and a disease characterized bypathophysiological hyper-vascularisation.
 15. The method according toclaim 14, wherein the inflammatory disease is selected form the groupconsisting of atherosclerosis, rheumatoid arthritis, asthma,inflammatory bowel disease, psoriasis, in particular psoriasis vulgaris,psoriasis capitis, psoriasis guttata, psoriasis inversa;neurodermatitis; ichtyosises; alopecia greata; alopecia totalis;alopecia subtotalis; alopecia universalis; alopecia diffusa; atopicdermatitis; lupus erythematodes of the skin; dermatomyositis of theskin; atopic eczema; morphea; scleroderma; alopecia greata Ophiasistype; androgenic alopecia; allergic dermatitis; irritative contactdermatitis; contact dermatitis; pemphigus vulgaris; pemphigus foliaceus;pemphigus vegetans; scarring mucous membrane pemphigoid; bullouspemphigoid; mucous membrane pemphigoid; dermatitis; dermatitisherpetiformis Duhring; urticaria; necrobiosis lipoidica; erythemanodosum; prurigo simplex; prurigo nodularis; prurigo acuta; linear IgAdermatosis; polymorphic light dermatosis; erythema solaris; exanthema ofthe skin; drug exanthema; purpura chronica progressiva; dihydroticeczema; eczema; fixed drug exanthema; photoallergic skin reaction; andperiorale dermatitis.
 16. The method according to claim 14, wherein thehyperproliferative disease is selected from the group consisting of atumor or cancer disease, precancerosis, dysplasia, histiocytosis, avascular proliferative disease and a virus-induced proliferativedisease.
 17. The method according to claim 16, wherein thehyperproliferative disease is a tumor or cancer disease selected fromthe group consisting of diffuse large B-cell lymphoma (DLBCL), T-celllymphomas or leukemias, e.g., cutaneous T-cell lymphoma (CTCL),noncutaneous peripheral T-cell lymphoma, lymphoma associated with humanT-cell lymphotrophic virus (HTLV), adult T-cell leukemia/lymphoma(ATLL), as well as acute lymphocytic leukemia, acute nonlymphocyticleukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronicmyelogenous leukemia, Hodgkin's disease, non-Hodgkin's lymphoma,myeloma, multiple myeloma, mesothelioma, childhood solid tumors, glioma,bone cancer and soft-tissue sarcomas, common solid tumors of adults suchas head and neck cancers (e.g., oral, laryngeal and esophageal),genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian,testicular, rectal, and colon), lung cancer (e.g., small cell carcinomaand non-small cell lung carcinoma, including squamous cell carcinoma andadenocarcinoma), breast cancer, pancreatic cancer, melanoma and otherskin cancers, basal cell carcinoma, metastatic skin carcinoma, squamouscell carcinoma of both ulcerating and papillary type, stomach cancer,brain cancer, liver cancer, adrenal cancer, kidney cancer, thyroidcancer, medullary carcinoma, osteosarcoma, soft-tissue sarcoma, Ewing'ssarcoma, veticulum cell sarcoma, and Kaposi's sarcoma, fibrosarcoma,myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma,angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma, mesothelioma, leiomyosarcoma,rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma,adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma,papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma,medullary carcinoma, bronchogenic carcinoma, seminoma, embryonalcarcinoma, Wilms' tumor, small cell lung carcinoma, epithelialcarcinoma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma,pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma,meningioma, neuroblastoma, retinoblastoma, glaucoma, hemangioma, heavychain disease and metastases.
 18. A method for treating ahyperproliferative disease or disorder, comprising the step of:administering a compound according to claim 1 or a pharmaceuticallyacceptable salt thereof to a patient prior to, during and/or after thepatient was subjected to at least one therapy selected from the groupconsisting of: radiation therapy; chemotherapy; immunotherapy;laser/microwave thermotherapy; and gene therapy using antisense DNAand/or RNA.